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Ageing Research Reviews 9 (2010) 117130

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Ageing Research Reviews


journal homepage: www.elsevier.com/locate/arr

Review

Role of chloroplasts and other plastids in ageing and death of plants and animals: A tale of Vishnu and Shiva
Wouter G. van Doorn a,*, Kohki Yoshimoto b
a b

Mann Laboratory, Department of Plant Sciences, University of California, Davis, CA 95616, USA Plant Immunity Research Team, RIKEN Plant Science Center, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan

A R T I C L E I N F O

A B S T R A C T

Article history: Received 24 May 2009 Received in revised form 25 August 2009 Accepted 25 August 2009 Keywords: Ageing Algae Apicoplast Bleaching Chloroplast Coral reef Death Degeneration Endosymbiosis Giant clam Kleptoplast Malaria Monocarpic Photosynthesis Plants Plasmodium Plastid Polycarpic Sea slug Survival Symbiosis Toxoplasma Vacuole

Chloroplasts (chlorophyll-containing plastids) and other plastids are found in all plants and many animals. They are crucial to the survival of plants and most of the animals that harbour them. An example of a non-photosynthesizing plastid in animals is the apicoplast in the malaria-causing Plasmodium species, which is required for survival of the parasite. Many animals (such as sea slugs, sponges, reef corals, and clams) consume prey containing chloroplasts, or feed on algae. Some of these incorporate the chloroplasts from their food, or whole algal cells, into their own cells. Other species from these groups place algal cells between their own cells. Reef-building corals often lose their intracellular algae as a result of environmental changes, resulting in coral bleaching and death. The sensitivity of the chloroplast internal membranes to temperature stress is one of the reasons for coral death. Chloroplasts can also be a causal factor in the processes leading to whole-plant death, as the knockout of a gene encoding a chloroplast protein delayed the yellowing that proceeds death in tobacco plants. It is concluded that chloroplasts and other plastids are essential to individual survival in many species, including animals, and that they also play a role in triggering death in some plant and animal species. 2009 Elsevier Ireland Ltd. All rights reserved.

1. Introduction The history of life knows of many momentous events. One was the invention of photosynthesis, the capacity to use the energy of sunlight and turn this into biochemical bonds. This evolutionary step apparently occurred in a lineage of prokaryotic cells. It resulted in a dramatic change in the earths atmosphere, as these photosynthetic prokaryotes produced oxygen.

* Corresponding author at: Mann Laboratory, Department of Plant Sciences, University of California, Davis, CA 95616, USA. E-mail address: wgvandoorn@ucdavis.edu (W.G. van Doorn). 1568-1637/$ see front matter 2009 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.arr.2009.08.003

Another enormous evolutionary event was the failed digestion, by a eukaryotic animal cell, of a prokaryotic photosynthetic cell. Most single-celled animals survive by consuming other cells. They capture their prey by engulng it. Once inside the animal cell, the prey cell becomes degraded by hydrolases. An evolutionary leap took place when the digestion of a photosynthetic cell became somehow impossible. A photosynthetically active prokaryote then remained inside the cytoplasm of the host eukaryote. Further evolutionary adaptations included the ongoing division of the guest prokaryote and the transfer of its offspring during cell divisions of the eukaryotic host (Keeling, 2004; Reyes-Prieto et al., 2007). When all of this was in place the eukaryotic plant cell was born, containing fully active chloroplasts (Greek khla ros = green; and plast = form, entity; related to plaste molder). The rst true s,

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plant cell and its multicellular descendants have been able to produce a considerable surplus, enough to sustain a relatively large biomass of non-photosynthetic organisms (animals, including mankind). Apart from being essential to survival, chloroplasts and other plastids have also been implicated in ageing and death. The term ageing will here be taken to indicate the deteriorative processses that increase the probability of death with increasing chronological age. Although ageing refers to what happens in individual multicellular organisms, the underlying deteriorative processes are often studied at the levels of tissues, cells, organelles, and molecules. Mitochondria, for example, have been have been implicated in ageing and are therefore studied in detail. Similarly, other organelles can be causal, or at least involved, in deterioration and death. The purpose of this paper is to give an overview of the current insights about the role of chloroplasts (photosynthetically active plastids) and other plastids in the survival and death plants and animals. Plastid is feminine of the Greek word plaste molder (from s, plastos, molded). Plastids will here be dened as organelles enveloped by a double membrane, which contain DNA, are capable of reproduction by ssion, are the site of manufacture and storage of important chemicals, and do not show the type of respiration found in mitochondria. In this review we will use the term chloroplast if referring to a plastid that carries out photosynthesis. Other plastids generally will be indicated either with their specic names or by the term non-photosynthesizing plastid. The term plastid will be used only when referring to the group of chloroplasts and non-photosynthesising plastids as a whole. Additionally, the term algae will be used for several organisms harbouring chloroplasts, including the blue green algae (phylum Cyanobacteria), green algae (Chlorophyta), diatoms (Heterokontophyta), and dinoagellates (Dinoagellata). Plastids are apparently found in most, if not all, living cells of higher plants. The Chlorophyll-containing, photosynthetically active chloroplasts are found in many plant cells. Non-chlorophyllous plastids are ubiquitous in cells of both photosynthetic and non-photosynthetic plant organs, for example in meristems, roots, the stem interior, leaves, and petals (Pyke, 2009). Similarly, many animal species harbour chrloroplasts or nonphotosynthesizing plastids, and depend on them. Non-photosynthesizing plastids are found in animals such as the malaria-causing Plasmodium. Many other animals contain either chloroplasts or chloroplasts in symbiotic algae. Examples are almost all reef-building polyps, sea anemones, and giant clams. It is thought that all coral reefs have formed because of the symbiosis between the polyps that produce the calcium carbonate skeletons, and photosynthetic algae in the phylum Dinoagellata. The algae in reef-building polyps provide up to 90% of their energy requirement (Marshall and Schuttenberg, 2006). The relevance of the symbiosis with algae is evident in the massive coral reef bleaching and subsequent death of the reef-building animals, which is due to degeneration and loss of the algae. It is thought that large parts of the coral reefs are currently under threat of disappearing (Lough and van Oppen, 2009; Pratchett et al., 2009). This paper will discuss, subsequently, endosymbiosis, the types of plastids found in plants and animals, their ultrastructure, their turnover, and their role in programmed cell death (PCD) and in the ageing and death of individuals. 2. Endosymbiosis The process of acquiring another cell that stays alive inside the cytoplasm is called endosymbiosis (Greek endo = inside, sym = together, biosis = living). The chloroplast endosymbiont in

plants apparently descends from cyanobacteria (Keeling, 2004; Reyes-Prieto et al., 2007) while another endosymbiont, the mitochondrion, derives from from a-proteobacteria (Gray et al., 2001). The prokaryotic nature of these endosymbionts is reected by their circular DNA, as well as their specic RNA and the full complement of the transcription and translation machinery. This machinery includes ribosomes and a typical set of enzymes involved in proper protein folding (class I chaperonins) which is only found in bacteria, chloroplasts and mitochondria (Fink, 1999). After entering their host cells early on in evolution, the endosymbionts lost most of their own genes. Many endosymbiont genes became translocated to the nucleus of the host cell, in a process called intracellular gene translocation. The activity of both the chloroplast and the mitochondrion thereby became under control of the host cell (Leon et al., 1998). The chloroplasts of green algae, red algae, blue green algae, and all multicellular plants are considered to be the result of primary endosymbiosis. This is the process whereby a photosynthetic cyanobacterium got enslaved by a eukaryotic animal cell (CavalierSmith, 2000). With the mitochondria also in place, this resulted in the rst eukaryotic algal cell. In other groups of organisms secondary endosymbiosis (or even tertiary symbiosis) has occurred. Secondary endosymbiosis is the acquisition of a eukaryotic algal cell, by a eukaryotic protozoan. After the secondary symbiosis, all organelles of the acquired algal cell disappeared, often with the exception of a remnant of the algal nucleus. Secondary endosymbiosis occurred in several taxa including the brown algae, diatoms, dinoagellates, and apicomplexans (Cavalier-Smith, 1999, 2000, 2002). Other types of symbiosis will also be described here, including the acquisition of chloroplasts from algae, by animals, and the acquisition of whole algal cells by animals, whereby the algal cells are placed inside the cells or between the cells. 3. Types of plastids in plants and animals Apart from chloroplasts (Fig. 1), plant cells can contain proplastids, a name often used for plastids that have not yet assumed a denite role. Proplastids are often found in meristematic cells. More differentiated non-chlorophyllous plastids serve a number of functions. The chromoplasts, for instance, contain high levels of carotenoids and other pigments, and provide colour. Storage plastids have been divided into amyloplasts (starch), elaioplasts (lipid) and proteinoplasts. Gerontoplasts (Greek geront = old man) are chloroplasts that are undergoing degenerative changes, in senescent tissues (Wise, 2007). Apicomplexans are unicellular animals, classied to a subgroup in the Protozoa. The apicomplexans include several diseasecausing parasites. Their plastid (apicoplast) still has some of its own genome and its own gene expression machinery, but also imports numerous proteins encoded by nuclear genes (Waller and McFadden, 2005; Obornk et al., 2009). For example, Theileria parasites cause death in cattle, Toxoplasma gondii (Fig. 2) causes fetal death in humans, and the malaria parasites (Plasmodium) kills many children and adults. The genus Plasmodium has over 200 species, at least 10 of which infect humans. Malaria is mainly caused by four species (Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale) that all infect human red blood cells, feed on them, and result in their destruction (Fig. 3). The worst type of malaria is caused by P. falciparum, which is kills approx. 12% of the people that get sick. Other Plasmodium species can infect various vertebrate animals. In contrast to the photosynthetically inactive apicoplast in Plasmodium and Toxoplasma, a similar, but photosynthetically active, plastid has been described recently in a protozoan. The host

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Fig. 2. Ultrastructure of Toxoplasma gondii. Am: amylopectin granule; Ce: centrioles; Co: conoid; Dg: electron-dense granule; Ga: Golgi adjunct (apicoplast); Go: Golgi complex; Im: inner membrane complex; Mi: mitochondrion; Mn: microneme; Nu: nucleus; Pm: plasmalemma; Rh: rhoptry. From Dubey et al. (1998).

Fig. 1. Chloroplast structure. (A) This chloroplast shows amyloplasts (starch bodies), indicated by a short arrow. The stroma is the uid inside the inner envelope. In the stroma lie stroma thylakoids (long thin arrows), which is an ER-like doublemembrane structure. These thylakoids are connected with the grana (long thick arrows) which are stacks of folded thylakoids. Picture from faculty.uca.edu/johnc/ cell1440.htm. (B) A chloroplast with small plastoglobuli (bodies containing mainly lipids and proteins), indicated with thin arrows. In other chloroplasts and in gerontoplasts these plastoglobuli can attain a consiserably larger diameter. Picture from http://botit.botany.wisc.edu/images/130/Photosynthesis/Chloroplast_EN.gif .

cell (genus Chromera) is apparently the closest known photosynthetic relative of the apicomplexan parasites (Moore et al., 2008). Kleptoplasty (Greek kleptein = to steal) is the retention of functional chloroplasts from algal prey. The chloroplast thus becomes stolen from a eukaryotic organism. This is in contrast to primary endosymbiosis, which involves the acquisition of a photosynthetic prokaryote. Remarkably, the algal cell is only

partially digested, sparing the chloroplast which stays intact and functional. The stolen chloroplasts are not carried into the progeny of the host (Clark et al., 1981). Kleptoplasty has been described in several dinoagellates, foraminifers, ciliates, and some sea slugs (molluscs). Dinoagellates are single-celled eukaryotes that use a agellum for locomotion. The kleptoplast in dinoagellates remains functional for a short period of time, usually not more than several days but in a few species up to 2 months. The kleptoplasts become digested and are replaced by chloroplasts from newly acquired prey (Gast et al., 2007; Minnhagen et al., 2008; Rumpho et al., 2008). Foraminifers (phylum Foraminifera) are small animals with pseudopods, which produce a shell with one or more chambers (Bernhard, 2003). Ciliates (phylum Ciliophora) are unicellular animals with motile hairs (Johnson et al., 2007). Fig. 4 shows examples of sea slugs that obtain chloroplasts from algae. Depending on the sea slug species, the chloroplasts are either stolen from green algae (Chlorophytae) or brown algae (Chromophytae). The kleptoplastic sea slug Elysia is shown in Fig. 4B. If it has eaten algae for only 2 weeks, it does no longer have to eat for the rest of its life, which is less than a year. The chloroplasts can function for up to 910 months. When the animals are deprived of algal food for a period of 8 months, their chloroplasts become more yellow, but the majority appeared still intact and with little ultrastructural change (Mujer et al., 1996;

Fig. 3. Plasmodium falciparium. (A) Plasmodium cells cause lysis of red blood cells. Picture from maib.mtandao-afrika.net/MAF060091/cause.htm. (B) Ultrastructure of Plasmodium. The apicoplast is indicated by and arrow. Black dots in the apicoplast are proteins visualized by immunogold detection. N: nucleus; FV: food vacuole (next to apicoplast). From Waller et al. (2000).

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Fig. 4. Some sea slugs run on solar power, after stealing chloroplasts. The animals depicted belong to the phylum Mollusca, class Gastropoda, order Sacoglossa. These slugs eat algae, digests all of the algal cell except the chloroplasts, and are able to use these to become photosynthetic themselves. (A) Placida dendritica showing the green network of ducts which contain green chloroplasts from its algal food. Picture from www.seaslugforum.net/factsheet.cfm?base=solarpow . Photo: Bill Rudman. (B) Elysia chlorotica, which has been fed algae. If young animals are fed algae for 2 weeks, they can survive for the rest of their life (about 810 months) without eating. Picture from www.3quarksdaily.com/3quarksdaily/2008/11/page/2/ . E. chlorotica lives in nature by feeding on the marine green alga Vaucheria litorea.

Rumpho et al., 2008). Chloroplasts in plants cannot survive for long without the expression of nuclear genes. It was a mystery, therefore, how the chloroplasts could remain active in the animal for so long, as there was no evidence that the sea slug also stole the algal nucleus. Surprisingly, it was found that at least one of the necessary algal nuclear genes had become incorporated into the nuclear DNA of the sea slug cells (Rumpho et al., 2008). A completely different method of acquiring photosynthetic capacity is the acquisition, by animals, of whole algal cells. An important distinction is the one between animals that have once in evolution acquired the algal cells, and transfer them into the progeny, and animals that have to obtain algal cells in each new generation. In the rst type of symbiosis with algal cells, the algae are passed on via maternal inheritance. At least one algal cell becomes incorporated in the egg, shortly after fertilisation. This type of symbiosis is found in freshwater polyps such as Hydra, several marine hydroids, corals, and sea anemones. Animals in this group can additionally obtain algae through feeding. For example, despite the transfer of Chlorella algae in their eggs, Hydra species can also take up Chlorella cells. The algal cells become endocytosed into single vacuoles, and are then transported form the gut side of the cells to the cell base (Trench, 1979). The second type of symbiosis in which the algal cells are involved depends on new acquisition in young indivials, which occurs through feeding (Trench, 1979). This type of symbiosis has been documented in phyla such Foraminifera, Radiolaria (amoeboid protozoa with mineral skeletons), Porifera (sponges), Cnidaria (hydrozoans, coral forming polyps, jellysh, and sea anemones), Platyhelminthes (atworms), and Mollusca (clams and sea slugs). The symbiotic algae are from phyla such as blue green algae, green algae, diatoms, and dinoagellates. The algae in this type of symbiosis are also referred to as zooxanthellae. This term results in confusion, however, because it has also been used to indicate symbiotic dinoagellates, or dinoagellates in the genus Symbiodinium (Trench, 1979; Venn et al., 2008). Although still widely employed, the term zooxanthellae will not be used here. Independent of the type of symbiosis (algae passed on from one generation to the next or acquired anew by each generation) the

algal cells are found either inside the animal cells, usually in vacuoles (intracellular symbiosis) or are placed between the animal cells (extracellular symbiosis). Intracellular or extracellular symbiosis is largely dependent on the algal species. For example, the green alga Chlorella is found as an intracellular symbiont in various freshwater animals (such as the sponge Spongilla and freshwater atworms such as Dalyella). The only exception to the rule, apparently, is the dinoagellate Symbiodinium, which is found as an intracellular and an extracelluar symbiont, depending on the animal species (Trench, 1979). Examples of intracellular symbioses are, apart from Chlorella, the green alga Tetraselmis which is found in marine atworms, the cyanobacterium Aphanocapsa in green algae, dinoagellates, and sponges (Carpenter and Foster, 2002), and the dinoagellate Amphidinium in atworms, mainly in the genus Amphiscolops (Taylor, 1978; McCoy and Balzer, 2004). The dinoagellate Symbiodinium is an intracellular symbiont in many reef forming corals (Trench, 1979; Gates et al., 1992) and sea anemones (Gates et al., 1992). Interestingly, some reef-building polyps also harbour intracellular nitrogen-xing blue green algae (cyanobacteria) in addition to the dinoagellate Symbiodinium (Lesser et al., 2004). The cyanobacteria were present inside a membrane. At least in some host cells the cyanobacteria were accompanied by Symbiodinium algae (Kvennefors and Roff, 2009). Examples of extracellular symbiosis with algal cells are also numerous. About 12 species of cyanobacteria have been found to show symbiosis with 38 genera of sponges (Carpenter and Foster, 2002). Cyanobacteria (usually Prochloron-related genera) are also found in ascidians, which are members of the family Ascidiacea in the phylum Chordata. These animals, also known as sea squirts, are sac-like marine lter feeders (Carpenter and Foster, 2002; Hirose et al., 2004). Diatoms include Licmophora, which shows symbiosis with the common marine atworm Symsagittifera (Convoluta) convuluta. Examples of dinoagellata are Scrippsiella in freeoating marine hydrozoans belonging to the phylum Cnidaria. The dinoagellate Symbiodinium is an extracellular symbiont in giant clams (Tridacna spp.; Trench, 1979). An example of a reef-building hydrozoan cnidarian with apparent extracellular algae is the soft coral Xenia (Fig. 5).

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Fig. 5. Photosynthetic cells in coral organisms. (A) A colony of the soft coral Xenia. The small thickenings along the oral tentacles are sacs lled with algal cells. Photo: Bill Rudman. From http://www.seaslugforum.net/factsheet.cfm?base=zoox2 . (B) Two oral tentacles showing the sacs with algal cells (red). Photo: Bill Rudman. From: http:// www.seaslugforum.net/factsheet.cfm?base=zoox2 . (C) Normal coral (left) and bleached coral (right). From sitemaker.umich.edu/section2group5/background.

Symbiodinium cells live in the oral tentacles which during the day are fully extended, capable of adsorbing sunlight (Fig. 5A). The algal cells are found in groups (Fig. 5B). Finally, lichens can also be considered to fall into this group, as they are a symbiotic unity of a fungal and a plant cell. This symbiosis has been reviewed by Honegger (1991). 4. Ultrastructure of chloroplasts and other plastids in plants and animals Chloroplast in plants are bound by two membranes, called outer and inner envelope. These membranes are separated by an intermembrane space. Inside the plastid is the stroma, an aequous uid (Fig. 1). The stroma contains many ER-like membranes called thylakoids. The thylakoid lumen is the name of the space between two thylakoids membranes. In many plants, the thylakoids fold to produce stacks of membranes, called grana (Fig. 1). The photosynthetic machinery is located both in the thylakoids and grana. Chloroplasts also contain droplets of osmiophilic material bound by a single sheet of lipids (a half membrane). These spherical structures are called plastoglobuli (Fig. 1B). They are the result of plastid membrane degradation and contain lipids and proteins (Brehelin et al., 2007). Starch grains are formed in the matrix (Fig. 1A) and are not bound by a membrane or a half membrane. Stromules (stroma-lled tubules) are mobile protrusions and interconnections between plant chloroplasts, usually 0.35 0.85 mm in diameter and of variable length. They form linear or branched structures up to 220 mm long (Natesan et al., 2005; Hanson and Sattarzadeh, 2008). These long, thin tubules contain no thylakoid membranes or chlorophyll. Stromules enable the transfer of molecules as large as ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; 560 kDa) between interconnected chloroplasts (Natesan et al., 2005; Hanson and Sattarzadeh, 2008). In addition to exchanging materials between chloroplasts, stromules may serve to increase the surface area of the chloroplast envelope for import and export, and anchor the chloroplast onto attachment points for proper positioning within the plant cell (Natesan et al., 2005; Hanson and Sattarzadeh, 2008). The ultrastructure of other plastids in plants has been discussed in detail in the reviews by Wise (2007) and Pyke (2009). Ultrastructural details have also been investigated in the apicoplasts in animals such as Plasmodium (Hopkins et al., 1999). The apicoplast is very small (Fig. 3) and often shows remarkably little structure in electron micrographs (Fig. 3B). The black dots observed in the apicoplast shown in Fig. 3B are due to immunogold staining, thus are not part of the normal structure as shown in electron micrographs. The apicoplasts are usually found to have a nely granular and sometimes brous texture. The ne

granules seem mainly plastid-type 70S ribosomes. Occasionally, some darker stained co-occurred with pools of rRNA. The brous nature of the apicoplast lumen has been suggested to represent the DNA (Waller and McFadden, 2005). The ultrastructure of the kleptoplasts in animals is almost the same as that of the chloroplasts in the algal cells from which they originate. For example, kleptoplasts have retained their outer and inner boundary membranes (Wise, 2007). Several examples of kleptoplast ultrastructure, in various animal species, are shown in the review by Rumpho et al. (2006). Trench et al. (1973) showed early work on the structure of algal cell chloroplasts in animals. Some other examples of ultrastructural studies on symbiotic algae include Amphidinium klebsii algae, both free-living (Blanco and Chapman, 1987) and in association with atworms (Mendes Lopes and Silveira, 2004). Symbiodinium has also been studied both in culture (Lesser and Shick, 1990) and in situ in animals such as jellysh (Wakeeld et al., 2000), coral reef ` cnidaria (Ladriere et al., 2008), sea anemones (Wakeeld et al., 2000; Muller-Parke et al., 2008) and giant clams (Bishop et al., 1976). In contrast to the chloroplast in multicellular plants, the chloroplast of many algal cells, both in species that are not symbiotic and those that are, contains a structure called the pyrenoid, which is involved in starch formation. Additionally, in the algal cells that are symbiotic the grana-type of thylakoid stacking is usually lacking, and there are few or no plastoglobuli. Other details are similar to the chloroplasts in multicellular plants, described above. Fig. 6 shows a Symbiodinium cell in a sea slug. Details of Symbiodinium in another sea slug species are found in Fig. 7. Note the presence of several chloroplasts, the pyrenoid body, a starch body, and a large lipid droplet in the algal cell. The presence of starch and lipids indicates that the algal cell produces surplus energy. 5. Interconversion, turnover, and degradation of chloroplasts and other plastids in plant cells The half-life of chloroplasts and other plastids, both in plants and animals, is as yet unknown. This is in contrast with mitochondria, which are known to become degraded and produced at a rate that is specic for each organ. Depending on the cell type the half-life of rat mitochondria varied between about 10 and 20 days (Menzies and Gold, 1971). Mitochondria apparently need to be degraded after a relatively short period, which might relate to avoiding the accumulation of a high mutational load in the mitochondrial genome. It has been suggested that the lack of histones in mitochondrial DNA accounts, at least in part, for the 10to 20-fold higher mutation rate of mtDNA compared to that in

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Fig. 6. The sea slug Pteraeolidia ianthina (phylum Molusca, class Gastropda, order Nudibranchia) harbours brown single-celled algae in its body, hence its colour. The algae are dinoagellates in the genus Symbiodinium. The slugs usually obtain the dinoagellate algae from cnidaria, such as sea anemones or soft corals, on which they feed. These cnidaria have previously incorporated the free-living algae. Picture from www.seaslugforum.net/factsheet.cfm?base=solarpow . (A) Whole animal. (B) A detail of the body. The brown lines across the body and down the cerata (appendices) are ducts lled with brown algal cells. (C) A section through a ceras to show the large numbers of algal cells. Bar = 40 mm. zoox: algal cells. Photos (AC): Bill Rudman. From http://www.seaslugforum.net/factsheet.cfm?base=zoox3 . (D) Scanning electron micrograph of a cluster of algal cells in a ceras. The darkest areas are the chloroplasts, which are mainly found appressed against the plasma membrane and the wall of the algal cells. Bar = 5 mm. Photo: Dianne Hughes. From http://www.seaslug.info/factsheet.cfm?base=zoox1 .

nuclear DNA. The mutation rate has been related, in turn, with errors in electron transport or in scavenging reactive oxygen species within the mitochondria (Kim et al., 2007). The same can possibly apply to chloroplasts, which might also be subject to a high mutation rate due to reactive oxygen species (ROS) and/or reactive nitrogen species (RNS) in case of aberrant changes in photosynthesis.

Plant plastids are highly interconvertible, sometimes with specic intermediate morphologies. Examples are the conversion of chloroplasts to chromoplasts during fruit ripening and ower petal development (Ljubesic et al., 1991; Cheung et al., 1993), and the conversion of amyloplasts to chloroplasts (Horner et al., 2007). During developmental PCD in leaves the chloroplasts become gradually dismantled. The organelles are

Fig. 7. Algal cells give the brown colour to the nudibranch Aeolidiopsis ransoni (phylum Mollusca, class Gastropoda). The algae are in the genus Symbiodinium, phylum Dinoagellata. (A) Whole animal. From http://seaslugsofhawaii.com/species/Aeolidiopsis-ransoni-a.html . (B) Section through a single algal cell with chloroplasts (c), a pyrenoid body (p), starch grains (s), and lipid body (lb). Bar = 1 mm. Photo (B): Dianne Hughes. From http://www.seaslug.info/factsheet.cfm?base=zoox1 .

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then called gerontoplasts. These gerontoplasts can become reverted to normal functional chloroplasts, even when they are already at an advanced stage of degradation (van Doorn, 2006, and references therein). The mechanisms for the degradation of materials in plant chloroplasts can be divided into at least four categories: (a) degradation of compounds inside the plastids without (or prior to) the plastids being transferred to a lytic vacuole, (b) transfer of materials out of the plastids, to be degraded in other organelles, (c) transfer of whole plastids into a lytic vacuole, where degradation occurs, and (d) degradation of the chloroplasts as a result of tonoplast rupture, whereby the remaining cell organelles are rapidly degraded by vacuolar hydrolases. We here present a short survey. Further details of chloroplast degradation have been reviewed recently by Sakamoto (2006), Hortensteiner and Lee (2007), Lim et al. (2007) and Krupinska (2007). 5.1. Degradation of compounds inside chloroplasts The chloroplasts leaf cells that are undergoing PCD become considerably smaller. This is accompanied by the complete loss of chlorophyll. Starch granule size also decreases (Inada et al., 1998; Matile, 2001). Small osmiophilic particles have been observed, associated with thylakoid degradation. They contain mainly free fatty acids, thylakoid lipids, and thylakoid proteins. These small particles might be the precursors of the much larger osmiophilic globules in the stroma, called plastoglobuli, which have been shown to contain similar compounds. Plastoglobuli in senescing leaves massively increase in volume and number (Krupinska, 2007; Brehelin et al., 2007). 5.2. Degradation of chloroplast components in other organelles Two types of globules have been seen to leave the chloroplasts during senescence. The rst type has been called Rubiscocontaining bodies (RCB). Apart from Rubisco (ribulose-1,5-biphosphate carboxylase oxygenase, the most abundant protein in photosynthesis) these globules contain other stromal proteins, for example glutamine synthase. The bodies are spherical (0.4 1.2 mm) and are bound by a double membrane. After being released to the cytoplasm, RCBs apparently become transported to a lytic vacuole by autophagy and are degraded there (Chiba et al., 2003; Ishida et al., 2008; Wada et al., 2009). In contrast to the relatively early release of RCBs, the release of considerably larger droplets occurs at a late stage of dismantling. These large droplets seem to be pushed out of the chloroplast. It has been suggested that they represent the plastoglobuli that are found to accumulate inside chloroplasts in senescing cells. It is unclear what happens with these droplets (Krupinska, 2007). Small senescence-associated vacuoles (SAVs) with intense proteolytic activity have been found in chloroplast-containing leaf cells of soybean, tobacco and Arabidopsis. The senescencespecic cysteine-protease SAG12 (senescence-associated gene 12) was localized only to SAVs (Otegui et al., 2005). In a tobacco line expressing green uorescent protein (GFP) targeted to chloroplasts, GFP was re-located to SAVs. Apart from GFP isolated SAVs also contained some chloroplast stromal proteins, including Rubisco and glutamine synthase, but lacked thylakoid proteins. These results indicate that the SAVs are involved in degradation of photosynthetic proteins of the chloroplast stroma (Martnez et al., 2008). The similarity between the composition of the RCBs (see above) and that of SAVs might indicate that RCBs fuse with SAVs or that RCBs become SAVs. The two types of vesicles might also be different, and both responsible for the degradation of stromal proteins outside chloroplasts.

5.3. Merging of chloroplasts with vacuoles; degradation inside vacuoles Degradation of cellular materials by the cell itself is called autophagy (Greek auto = self; phagein = to eat, consume). Microautophagy is the uptake of materials, sometimes organelles, by a lytic vacuole. It usually occurs by the invagination or protrusion of the tonoplast (vacuolar membrane) to engulf cytoplasmic materials. Macro-autophagy is carried out by a unique organelle, which has been described in detail both in animals and yeast. A portion of cytoplasm, which can include various organelles, is surrounded by a growing double-membrane structure, the isolation membrane. Upon closure of the double membrane, the structure with its enclosed cytoplasmic material is referred to as the autophagosome (Mizushima, 2005). In animals the outer membrane of the autophagosome subsequently fuses with a lysosome, a small vesicle that contains hydrolases. The autophagosome is then called autolysosome. The hydrolases rst degrade the inner membrane of the autophagosome and then the engulfed material (Mizushima, 2007; Xie and Klionsky, 2007). In yeasts the autophagosome rather fuses with a large lytic vacuole, which also results in degradation of the material engulfed by the autophagosome (Baba et al., 1994). Two early studies suggested that chloroplasts merged with or were taken up by large vacuoles during leaf senescence (Peoples et al., 1980; Wittenbach et al., 1982). Similarly, the results of Minamikawa et al. (2001) indicated initial degradation of Phaseolus vulgaris leaf chloroplasts in the cytoplasm and later presence of these chloroplasts inside the vacuole. Some recent papers shed further light on the presence of chloroplasts in lytic vacuoles and the possible role of autophagy in the process. Wada et al. (2009) individually darkened leaves of Arabidopsis plants, which promoted their yellowing. The time to yellowing was almost the same in wild-type and in an autophagydefective T-DNA insertion double mutant in the autophagy-related (ATG) genes 4a and 4b (atg4a4b-1). The number and size of chloroplasts decreased in darkened leaves of wild-type plants, but remained about constant in the atg4a4b-1 mutant. Chloroplasts were found in the wild-type vacuole, but were not present in the vacuoles of the autophagy-defective mutant. These data show that chloroplast transfer to the vacuolar lumen depends on autophagyrelated genes. Autophagy genes in plants can be involved both in micro- and macro-autophagy. Effects of knockdown or knockout of autophagy genes, therefore, is good evidence for a role of autophagy, but it does not tell whether it is micro- or macroautophagy. No evidence has been given, thus far, for the uptake of whole chloroplasts by vacuoles through micro-autophagy, but this mechanism cannot be excluded. If macro-autophagy in plants is dened as in yeast and animals (by the presence of autophagosomes and autolysosomes), a role of macro-autophagy in the degradation of chloroplasts is also unlikely. Strictly speaking, no autophagosomes have been shown in intact plants. The structures that have been shown are very similar to autophagosomes, but were already acidic and already contained hydrolases before they surrounded and then degraded a portion of the cytoplasm. They should therefore be called lytic vacuoles, or autolysosomes. Such autolysosomes have been shown in root tip meristem cells in several plants (Buvat, 1971; Buvat and Robert, 1979; Marty, 1978, 1999). Others have also claimed evidence for the presence of similar autophagosomes/autolysosomes in other cells of intact plants (for example Liu et al., 2005) but these claims are relatively weak as they were not backed up by showing engulfment of a portion of the cytoplasm and the presence of hydrolases within the vesicle. Recent molecular studies by using ATG8 protein, a purported marker protein for autophagosomes, have also been taken to suggest the presence of autophagosomes in intact plants

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(Yoshimoto et al., 2004; Contento et al., 2005; Thompson et al., 2005). However, this evidence is similarly still putative. Moreover, the putative autophagosomes as visualized by ATG8 tagged with green uorescent protein seemed not large enough to engulf a whole chloroplast. The available data thus show autophagosomes/ autolysosomes in a few types of cells of intact plants but do not provide good evidence for such structures in cells containing chloroplasts. The transfer of whole chloroplasts into autophagosomes/autolysosomes, where they would become degraded or become transported to a large lytic vacuole, seems therefore a remote possibility. The best evidence thus far for the transfer of chloroplasts to vacuoles is neither through micro- nor through macro-autophagy. Peoples et al. (1980) showed (in their Fig. 9) that the tonoplast of a large vacuole merged with the outer envelope of a chloroplast. Once this merging is complete the hydrolases resident in the vacuole will be able to destroy the inner chloroplast membrane and the remainder of the organelle. This is similar to what happens with autophagosomes in yeast, which release their inner vesicle, and the contents of that vesicle, into a large lytic vacuole (Baba et al., 1994). Taken together, the data indicate that transfer of chloroplasts to lytic vacuoles can occur through merging of the outer envelope of the chloroplast with the tonoplast. Although autophagy genes are involved in chloroplast to vacuole transfer, their precise role remains to be elucidated. 5.4. Chloroplast degradation after tonoplast rupture A fourth type of chloroplast degradation during PCD in plants has been found in leaf mesophyll cells that are induced to produce dead xylem tracheid elements (TEs). Little internal degradation was observed in the mesophyll cells chloroplasts, until the very end of the PCD process. Death came about by the rupture of the vacuolar membrane (tonoplast). This resulted in the release of the hydrolases that had accumulated inside the vacuole. Upon release, these hydrolases degrade the chloroplasts and the other remaining organelles in a very short period (Fukuda et al., 1998; Kuriyama, 1999). Generally, programmed cell death in plant cells is directly due to tonoplast rupture (van Doorn and Woltering, 2005), but most chloroplasts in vivo seem to become degraded before such rupture takes place. For example, Inada et al. (1998) found that in leaves attached to whole rice plants the number of chloroplasts per cell became close to zero before tonoplast rupture. This means that the chloroplast destruction in the TE system might not be representative of most chloroplast-containging cells in vivo. The data in this section suggest that there may be several ways of chloroplast degradation in plants. The data indicate that the mechanisms are not mutually exclusive, as chloroplast degradation can proceed until little is left of the organelle, while it is still residing in the cytoplasma, while transfer of apparently rather intact chloroplast to the vacuole has also been reported during leaf senescence. 6. Turnover and degradation of chloroplasts and other plastids in animal cells Little work has been reported dealing with the fate of the kleptoplast. In the planktonic ciliate Strombidium capitatum, when fed algae for the rst time, chloroplasts were initially still in intact algal cells. These algal cells were localized in vacuoles of the host. Rapidly thereafter, the chloroplasts (kleptoplasts) were found free in the ciliate cytoplasm (Stoecker and Silver, 1990). Chloroplasts that were stolen from algae by sea slugs such as Elysia viridis and Elysia (Tridachia) crispata were initially present in digestive cells, in a vacuole. The vacuolar membrane soon disappeared, thus the organelle became part of the cytoplasm (Trench, 1979). These data

suggest that algae are transported to vacuoles where the algal cell becomes degraded while the chloroplast remains. This is followed by dissolution of the vacuolar membrane. Some of the cytoplasma-localised chloroplasts were later on engulfed by autophagosomes. This was followed by fusion of the autophagosome with a small lysosome. The lysosomal hydrolytic enzymes degraded the chloroplast (Trench, 1979). This process of chloroplast turnover in animal cells is remarkably similar to that of mitochondrial turnover. It has been thought that lysosomal degradation occurs as soon as the chloroplasts become defunct (Trench, 1979). The kleptoplast, when degraded in autolysosomes, serve as food. When not yet residing in animal tissue, the algae involved in symbiosis with animals are usually oating freely in the water. They are picked up by feeding, after which they become properly ensconced. The reproduction process of the symbiotic algae is apparently mainly asexual, i.e. they divide into two identical individuals. Chloroplasts in algal cells, it might be hypothesized, can become degraded in a way similar to that found in cell of higher plants (see Section 5). Whole algal cells inside animal cells might become degraded inside the vacuole where they are normally found, or in other ways. Algal cells that are localized between host cells might be released or become ingested by phagocytic activity. Under optimal light conditions, the colonial radiolarian species Collozoum interne was found to ingest their symbiotic dinoagellate algae at a rate that corresponded with algal reproduction. When the animals were placed under low light the net number of algae (the product of algal reproduction and algal destruction) decreased, in association with sequestering and degradation of the algae in lysosomes (Anderson, 1983). The dinoagellate Symbiodinium occurs in a symbiotic association with sea slugs such as Melibe sp. A large proportion of the fecal material of the animal consisted of degenerate symbiotic algal cells. It was thought possible, therefore, that these sea slugs obtain part of their nutrients by digestion of some of their algal symbionts (Kempf, 1984). It was observed that the green hydra, Chlorohydra viridissima, kept under natural conditions, actively expelled their symbiotic algal cells during predatory feeding on Artemia (brine shrimp, a small crustacean). Expulsion occurred from the endodermal digestive cells (Fishman et al., 2008). One mechanism of getting rid of excess symbiotic algal cells, therefore, is simple expulsion. The symbiosis with algal cells in reef-building corals is very sensitive to environmental factors, whereby the algal cells can become degraded in situ. Algal cells, normally present in non-lytic vacuoles of host cells can also leave the organism when still inside the host cells. This occurs when these host cells lose their adherence to other host cells (Gates et al., 1992). The degradation or disappearance of algal cells in coral reef species often results in early death of the individuals. This will be discussed in more detail in Section 8. The turnover rate of the non-photosynthesizing apicoplast is apparently not known, and it has apparently also not been reported how the apicoplast becomes degraded. 7. Role of chloroplasts and other plastids in plant survival, plant programmed cell death, and death of whole plants The importance of chloroplasts for plant survival is obvious. Chloroplast produce sugars, thus energy. They also are a source of amino acids and glycolipids. Chloroplasts are also the primary locus for the synthesis of fatty acids in green plants (CavalierSmith, 2000) and are apparently the sole locus for the biosynthesis of the thiazole moiety of vitamin B1 (Julliard and Douce, 1991). Moreover, chloroplast produce isoprenoids, and convert these to two major groups of lipid-soluble antioxidants in photosynthetic

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tissues: vitamin E (phylloquinone) and carotenoids (Schultz et al., 1985; DellaPenna and Pogson, 2006). Other isoprenoids are required for the synthesis of compounds that participate in respiration (ubiquinone), photosynthesis (carotenoids, chlorophylls, plastoquinone) and regulation of plant growth and development (cytokinins, brassinosteroids, gibberellins, abscisic acid). Many plant isoprenoids, additionally, are secondary metabolites that function in protecting plants against herbivores and pathogens (Phillips et al., 2008). Chloroplasts thus not only capture energy from sunlight and converts this into carbohydrates, but also produce many important or essential chemicals. When discussing the role of chloroplasts in plant survival and whole-plant death, it is imperative to dene a few terms. In animals, the term ageing usually indicates the deteriorative process that increases the probability of death with increasing chronological age. The terms senescence is often taken to be a synonym of ageing. In plants, the terms aging and senescence have also been used for the same purpose, but both terms contain ambiguities. The term aging is also used to indicate any change with age. The term senescence in plants not only refers to whole individuals but also the processes that precede the death of an organ (such as a leaf) in an otherwise healthy individual, whereby the death of such an organ is often promoting individual survival. In order to avoid confusion, the term senescence of whole plants will here be used when indicating the process that is equivalent to ageing in animals. Many species of multicellular land plants die after one growth season (annual plants), after two seasons (biannual plants) or after more seasons (perennial plants). These differences are partially overlapping with another distinction. In many plant species death quickly follows seed production (in monocarpic plants) or takes place only after many seasons of seed production (polycarpic plants). Annual and biannual owering plants usually die after they have set seed, thus are monocarpic. Perennial owering plants can also be monocarpic, as in bamboo which lives for 5070 years, depending on the species, set seed and die. Other perennial plants die after several seasons of seed production, thus are polycarpic. Their death is much less visible as in monocarpic plants, which usually become completely yellow due to chloroplast degradation. Two types of whole-plant death, thus, need to be explained: the ones due to monocarpic and polycarpic whole-plant senescence. Although investigated in only a few species detail, the death of many monocarpic plants is controlled by the reproductive structures, as removal of the inorescences or seeds prevent death. Death of monocarpic plants is preceded by cessation of growth and massive leaf yellowing. In soybean plants, for example, growth of the leaves ceased at the onset of owering. Relatively immature seeds were found to secrete a chemical that induced leaf yellowing and death of the leaf cells. Removal of young seeds prevented this yellowing and the subsequent death of the whole plant. Young seed removal did not reinstall growth of the aboveground parts of the plant (Nooden and Penney, 2001). Leaf yellowing is due to chlorophyll degradation in chloroplasts. It is a symptom of the retrieval of valuable compounds from the plant to the growing seeds. Details of chloroplast degradation have been discussed in Section 5. These data indicate that chloroplasts are at least passively involved in monocarpic senescence. Interestingly, chloroplasts have also been found to actively promote the symptoms that precede death. In intact tobacco plants, Zapata et al. (2005) reported that the knockout of the chloroplast gene ndhF plants resulted in a considerable (30 days) delay in visible leaf yellowing. The transgenic plants did not show a delay in the onset of owering. Rather, owering in transgenic plants occurred about a week earlier than in wild-type plants. The onset of owering in wild-type tobacco was accompanied by the onset of leaf yellowing (progressing form basal to apical leaves) of

the branches that support the reproductive structures. However, the leaves of the transgenic plants did not show a similar degree of yellowing until the late stages of fruit development, around 40 days after the onset of owering. These data indicate that the death-inducing effect of the reproductive organs, typical for monocarpic plants, acts through chloroplasts. The mechanism of action is unknown but might include the production of excess free radicals in chloroplasts (Martn et al., 2004). It might be assumed that delayed leaf yellowing also delayed the time of whole-plant death, but this was not reported in detail. The cause of death of plants that set seed many times (polycarpic senescence) is still largely unknown. Deterioration in the meristems of these plants is apparently not the reason of death. One of the reasons for death is that the size of the plant becomes too big, which causes problems in water relations. This in turn might explain the changes found in hormones concentrations, the closure of the stomates, and the decrease in the rate of photosynthesis. Water stress usually results in increased abscisic acid (ABA) levels. The high ABA levels, in turn, can explain stomatal closure and the decrease in photosynthesis. However, even in the absence of apparent water stress, ABA levels can be increased in old plants (Munne-Bosch, 2007). Additionally, Seo et al. (2000) reported that DS9, a protein localised to chloroplasts, regulated the programmed cell death which is a response of tobacco mosaic virus infection in tobacco leaves. This type of cell death is very local, and close to the point of entry of the pathogen. The dead cells stem the advance of the pathogen in the plant. This type of plant PCD, therefore, delays death of the whole plant rather than induces it. Taken together the data in this section show that chloroplasts are at least passively involved in the death of monocarpic whole plants. In tobacco the chloroplasts also play an active role as they hastened the leaf yellowing that proceeds whole-plant death. Polycarpic senescence also involves a deterioration of chloroplast functioning, which seems a result of other physiological changes. No active role of chloroplast in this type of whole-plant death has yet been established. 8. Role of chloroplasts and other plastids in the survival and death of animals The discussion in this section will rst focus on the two main types of plastids in animals, and on the symbiosis with algal cells. Secondly, a few details of the role of chloroplasts in the death of coral-reef-building animals will be reviewed. 8.1. Apicoplasts In the parasite T. gondii (Fig. 2) the apicoplast-localized fatty acid synthesis (FAS II) pathway is different from that in the host cell. The FAS II pathway has been found to be required for parasite survival (Mazumdar et al., 2006). The apicoplast is the only site of de novo synthesis of lipoate, a salt or ester of lipoic acid (6,8dithiooctanoic acid). Lipoate is required for lipoylation thus for the proper functioning of several proteins, for example some enzymes involved in energy metabolism, amino acid degradation and folate metabolism. T. gondii cannot produce lipoate in organelles other than the apicoplast, as it has lost the mitochondrial lipoate synthase gene (Crawford et al., 2006). Similarly, Gunther et al. (2009) found evidence for the idea that lipoate biosynthesis is specic for the apicoplast of Plasmodium, and that the apicoplast is required for protein lipoylation. Some antimalarial drugs mainly target the apicoplast. They often cause modest antimalarial effects initially but are especially potent against the progeny of treated parasites. The daughter cells of the host inherit nonfunctional apicoplasts and then fail to complete their devel-

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opment in red blood cells (Dahl and Rosenthal, 2007, 2008; Goodman et al., 2007). However, a direct effect on Plasmodium survival has also been found by targeting the apicoplast. Ramya et al. (2007) reported that several inhibitors of the fatty acid synthesis pathway displayed a direct and striking killing effect in Plasmodium falciparium. Similarly effective was succinyl acetone, an inhibitor of heme biosynthesis in the apicoplast. These data strongly support the idea that the apicoplast is necessary for the normal life cycle of the parasites. 8.2. Kleptoplasts When evaluating the role of stolen chloroplasts or symbiotic algae, it must be kept in mind that the organelles and cells can always serve two purposes: (a) production of energy and materials through photosynthesis, and (b) production of energy and materials after being digested by the host. The contributions of each of these two sources of food are quite hard to disentangle. As noted, a planktonic ciliate (S. capitatum) steals functional chloroplasts from ingested algal cells. If ciliates that had already obtained chloroplasts were held in the light but given no more algae, the individuals decreased in size and number. The number of chloroplasts per individual decreased, but the starving ciliate cells still contained a few chloroplasts at the end of their lifespan (40 h or more). Similarly, the ciliate population did not increase when the animals could only feed on prey that lacked chloroplasts (Stoecker and Silver, 1990). These data suggest that survival of these animals depended on almost continuous restocking with chloroplasts. The relative role of photosynthesis and chloroplast digestion in the survival rate was not separated in these tests. The marine dinoagellate Dinophysis caudata can only survive if it feeds on the ciliate Myrionecta rubra (=Mesodinium rubrum), from which it steals chloroplasts. A 3-month starvation experiment showed that D. caudata remained photosynthetically active for about 2 months when not supplied with ciliate prey, and had by then lost their chloroplasts. When the normal prey was available again, the D. caudata cells were rapidly acquired plastids and resumed photosynthetic activity (Park et al., 2008). As in the ciliates discussed above, these animals, therefore, cannot survive prolonged periods of absence of their ciliate prey, i.e. absence of new chloroplasts. Similarly, stolen chloroplasts contributed to the survival of sea slugs during times of food shortage. When food was not scarce, there was only 510% transfer of 14C xed by chloroplasts to the remainder of the animal, suggesting that the animal could do without the chloroplasts under such conditions. However, when food was scarce, the chloroplasts were important for survival. For example, adult individuals of well-fed Elysia timida that were withheld food and kept in the dark for 18 days still showed no difference in death rate compared with those kept in the light, but by day 28 those kept in darkness showed a decrease in size and up to 30% lower survival rate, compared with those held in the light (Gimenez Casalduero and Muniain, 2008). The sea slug Elysia chlorotica is also capable of surviving without food for a long time, after eating algae, providing that they are in the light. The animals are hermaphrodytes that die synchronously in the late spring of each year, after laying egg masses. Electron microscopy of the animal cells that were undergoing PCD prior to death showed DNA fragmentation, chromatin condensation and formation of primary lysosomes. This was followed by fragmentation of the nucleus. Together with all other organelles in the cells the chloroplasts degenerated until none were left. The cytoplasm formed a large central autolysosome, which contained numerous viral particles. Viruses were abundant at the end of the life cycle of the animal. The coincidence of viruses and death suggested a possible causal relationship between the two, whereby loss of resistance to

viruses might induce death (Mondy and Pierce, 2003). The timing of death in this slug species, therefore, seemed independent of the presence of stolen chloroplasts, and was related to the time of reproduction. 8.3. Algal cells Several species of radiolarians and foraminiferans, containing symbotic algae, are common in surface water and in open-ocean plankton. Generally, they also feed on planktonic species that contain no chloroplasts and are therefore not fully dependent on photosynthesis. This strategy is called mixotrophy. The positive role of the symbiotic algal cells in the survival of the individuals in these species increased during periods of starvation (Michaels and Silver, 1988). In an experiment with the green freshwater polyp Hydra (Chlorohydra) viridissima (class Hydrozoa, phylum Cnidaria) the animals were fed with the green alga Chlorella. Subsequently, they were kept without algae. The survival rate of 12-day starved animals was highly dependent on the number of photosynthetically active algal cells that were left in their bodies. A sharp decrease in survival rate was correlated with a number of algal cells that was less than 30% of original (Muscatine and Lenhoff, 1965). The algal symbiont, therefore, is clearly important for survival in times of starvation. In another experiment Hydra viridis with and without symbotic algal cells (Chlorella) in their body were reared under moderate starvation. Asexual growth (the number of new polyps) was drastically reduced in starving animals bearing no algae. Sexual reproduction was also affected by the presence of the symbionts. In well-fed animals no female or male reproductive parts were found. However, in moderately starved animals female gonads were produced only in animals having symbiotic algae. In contrast, the formation of male reproductive was independent of the presence of algae (Habetha et al., 2003). Hydra has been considered to be an example of a multicellular organism that does not show ageing, i.e. it does not show increased mortality with chronological age. The evidence for this idea stems from experiments of Martnez (1997). No increase in mortality was found in a study whereby Hydra vulgaris individuals were kept for 4 years. It might be argued that this time is rather short to show absence of ageing. The alternative hypothesis is that the animals have a long lifespan. Martnez (1997) thought this hypothesis to be less plausible, based on the prediction of evolutionary theory that there will be a positive correlation between age of rst reproduction and maximum lifespan, so that animals that start reproducing soon after birth have shorter lifespans. Indeed, when taking 4 years to be its maximum longevity, Hydra turned out to be an exception among many other species, when plotting the age of rst reproduction against maximum longevity. However, we take it that these data are not convincing enough to support the claim of lack of ageing in Hydra. Still, the long life of Hydra seems related to the extensive capacity to regenerate and self-renew. This capacity is due to the presence of three distinct stem cell lineages that are present throughout the body: ectodermal and endodermal epithelial stem cells, and interstitial stem cells (Bosch, 2008). Small photosynthetic atworms in plankton contain symbiotic algae, but also eat organisms without chloroplasts (Stoecker et al., 1989). The benthic atworm Amphiscolops langerhansi has been found to be unable to achieve sexual maturity without its symbiotic algae (Taylor, 1971), and the closely related atworm Convoluta roscoffensis was completely dependent on its symbiotic algae for growth (Provasoli et al., 1968). The role of the algae in ageing is apparently unknown. Many jellysh are also potentially autotrophic (i.e. they do not need to eat if in adequate light) because of their symbiosis with algae (Kikinger, 2008). The jellysh Mastigias papua (Sugiura,

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Fig. 8. Some giant clams (phylum Mollusca, class Bivalvia, fam. Tridacnidae) completely or partially run on solar power, after establishing a symbiosis with algal cells that live in their tissues. Individuals in some of these clam species can live for more than 150 years. Whether the presence of the algae is the reason for their long life-span is at present not known. (A) Tridacna crocea, the burrowing giant clam. From http://www.reefcentral.com/forums/showthread.php?threadid=1072234 . See for the wide range of colours in this species: http://reefkeeping.com/issues/2007-10/jf/index.php . The species is among the smallest of the giant clams. (B) Another species in the genus, probably Tridacna gigas. If so, it can become about 1.3 m long. From www.itsnature.org .

1964) and the upside down jellysh Cassiopeia andromeda (Ludwig, 1969) were found to grow property only if infected with algal cells. Effects of the symbiotic algae on mortality at various ages have apparently not been reported. Algal cells are also essential to the survival of individuals in coral-building species. The animals, at any age, are unable to survive when they are withheld to acquire algal cells (Yakovleva et al., 2009). More details on the role of algae in the survival and death in these species are discussed below. In several species of giant clam (genus Tridacna, Fig. 8) the young individuals ingest various strains of the dinoagellate Symbiodinium. Young Tridacna squamosa fed with algal cells had higher survival rates than the controls. Even when no food was given (the environment was millipore-ltered seawater) the juveniles that contained algal cells survived and grew well, for over 10 months, when held in white light as the sole energy source (Fitt and Trench, 1981). So the role of the symbiotic algae in juvenile survival is considerable, at least under conditions of starvation. Aquarium enthusiasts know that giant clams need considerable light levels for survival. According to their experience Tridacna crocea and Tridacna maxima require the most light, Tridacna squamosa needs less, while Tridacna deresa is probably the most forgiving in terms of light, compared with the other species mentioned (reef central page; http://www.reefcentral.com/forums/showthread.php). Indeed, Tridacna maxima mostly occurs in shallow waters, while Tridacna squamosa may occur down to greater depths. Jantzen et al. (2008) indicated that this difference in vertical distribution could only partly be attributed to differences in their photosynthetic performance. Both species showed the same rates of photosynthesis at relatively low light levels, but Tridacna squamosa had lower rates at high light levels, compared with Tridacna maxima. These data refer to rapid changes in the light environment, thus exclude long-term adaptations. It has been suggested that giant clams can live for more than 150 years, while some individuals become over 200 years old. It also has been proposed that the giant clams can reach their size because of the photosynthates produced by their algal symbionts, and that their long life is also due to the algae (http:// www.itsnature.org/sea/other/giant-clam/). However, these claims seem not yet independently tested. 8.4. Algal cells and coral bleaching As a result of high environmental stress or pathogen infection, the coral animals often lose their algal cells either by expulsion or

through death and digestion. The visible result is coral bleaching (Fig. 5C). The animals on the right side of Fig. 5C have lost their pigment but have apparently not yet died. Exposure to air during low tides and damage from solar radiation in shallow water environments are two of the environmental reasons for coral bleaching. Changes in water temperature are also an important cause. A temperature rise as little as 12 8C for 510 weeks (or a 3 5 8C decline for 510 days) are adequate to induce coral bleaching. Global warming results in an increase in surface water temperature but also causes hurricanes, which by themselves lead to coral bleaching (Lough and van Oppen, 2009). Coral bleaching was accompanied by a reduction in algal density between 50 and 90%, depending on the species. The reasons for this decrease include (a) degeneration of algal cells in situ, (b) release of algal cells, and (c) release of host cells containing ` algae (Brown et al., 1995). Ladriere et al. (2008) conrmed that the algal cells in bleached coral cnidarians exhibited various stages of disorganization. Several algal cells had become vacuolated. Some of the algal cells were still associated with the vacuoles of the host cells, but others were already free. Gates et al. (1992) corroborated that the bleaching of the reef coral Pocillopora damicornis was accompanied with detachment of host endoderm cells that still contained the algae in their vacuole. Once the host cells became detached and were released to the seawater, they disintegrated, thus the algae were free. One of the causes of coral bleaching, therefore, seems a dysfunction in cell adhesion. The physiological reasons for coral bleaching are as yet largely unknown. Interestingly, Tchernov et al. (2004) found evidence for the chloroplast thylakoid membranes in the alga as one of the key determinants of thermal stress sensitivity. The critical threshold temperature separating thermally tolerant from thermally sensitive strains of algae was determined by the degree of thylakoid lipid saturation. Additionally, high irradiance in the algal cells resulted in accumulation of high levels of reactive oxygen species, which triggered death and expulsion of the algae from the host. Thermal bleaching also has been related to photoinhibition in the chloroplast. Photoinhibition is due to oxidative stress, i.e. a surplus of reactive oxygen species at photosystem II. Photoinhibition takes place only when the rate of photodamage to photosystem II exceeds the rate of repair. A moderate increase in temperature was found to accelerate photoinhibition primarily through inhibition of the repair process (Baird et al., 2009; Baker et al., 2009). Photoinhibition after an increase in water temperature was found to be dependent on the clade of Symbiodinium cells. Differences in sensitivity among coral species therefore may be determined, at least in some species, by the

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symbiont (Baird et al., 2009). However, a relationship between Symbiodinium clade and bleaching was not always found. Coral species vary considerably in their response to thermal stress. Some common Symbiodinium clades, such as C1 and C3, have been found to occur in host species with a very divergent response to temperature, including the highly susceptible Seriatopora hystrix, as well as in several species that are known to be quite resistant to thermal stress, including those in the genera Favia, Goniastrea and Platygyra (Baird et al., 2009). Depending on numerous factors, the coral reef animals can sometimes recover from bleaching (Pratchett et al., 2009). Recovery from a natural bleaching event in various reef coral species was rather slow (more than 20 weeks) and apparently due to division of the remaining algal cells. The rate of recovery was hampered by the continual loss of apparently healthy algal cells from the bleached coral (Jones and Yellowlees, 1997). Bleaching has also been studied in sea anemones, where the cell biological processes seem similar to those in reef-building cnidar` ians. This follows of the ndings of Gates et al. (1992) and Ladriere et al. (2008), for example, who studied Aiptasia pulchella in addition to their investigation of several reef-building species (see above). Taken together, the data in this paragraph indicate that the nonphotosynthesizing apicoplast is essential to the survival of desease-causing apicomplexans. Chloroplasts are essential for survival, at least during food shortage, of several organisms that steal chloroplasts and insert these inside in their cells. Moreover, algal symbionts are very important in many organisms that harbour them. Quite little is as yet known about the details of role of chloroplasts on ageing in these animal species. Only in some reef-building species, it has been suggested that death is due to the chloroplasts, whereby membrane properties and free radicals that inhibit photosystem II have been implicated. 9. Conclusions In Hinduism mythology, Brahma is the creator, Vishnu the maintainer, and Shiva the destroyer and recycler. The chloroplast is a guest organelle that maintains almost all life on earth (some bacteria are the exception as they are able to live on the reduction of inorganic compounds). The chloroplast therefore has a high Vishnu character. Chloroplasts maintain their life through the cycle of birth (chloroplast ssion) and death (chloroplast degradation). Chloroplast, therefore, do not escape the reign of Shiva. But remarkably, chloroplasts are able to do a dance between Vishnu and Shiva. In a cell undergoing PCD, they can degenerate to a large extent but upon the right stimuli they can reverse this process of internal degradation, whereby they re-synthesize the structures necessary for normal photosynthesis. This reversal is associated with a reversal of PCD in the cell that harbors the chloroplasts involved, resulting in a long life of the cell thereafter. Apart from this dance between maintenance and death, chloroplasts can also be actively involved in causing death in plant cells, so have also Shiva character. Plant survival depends on active chloroplasts and other plastids. The death of plants in species that produce seeds only once (monocarpic whole-plant senescence) is due to the reproductive structures and is closely related to chloroplast degradation. In tobacco the death-signal of the reproductive structures was found to be transmitted through the chloroplasts. A passive or active role of chloroplasts is much less obvious in the death of plants that produce seeds during several growth seasons (polycarpic whole-plant senescence), although a decrease in chloroplast function has been implicated. As in plants, the survival of several animals depends on nonphotosynthetic plastids or on chloroplasts. The non-photosynthesizing plastid in the single-celled animals that cause diseases such as malaria have been found to be essential to survival, as plastid-

directed toxic chemicals were effective in killing these cells. Individuals of species in which the animals are withheld the opportunity to obtain chloroplasts or whole algal cells live shorter than those that have obtained the organelles or cells, especially in times of starvation. Nonetheless the precise role of chloroplasts in ageing and death is still largely unknown. As an example, the algal cells that live in symbiosis with reef-building animals are essential for colony survival. Coral bleaching (loss of algal pigment) is the result of death and/or release of the algal cells. In some examples death was mediated by the chloroplasts. This type of death, however, is induced by an external factor. It is not known, apparently, whether the sensitivity of the reef-building animals to bleaching increases with age. Additionally, the role of the algae in the ageing of these animals, when grown under non-stress conditions, seems as yet unknown. This review, therefore, points to many lacunae in our knowledge about the role of chloroplasts (and other plastids) in ageing and death. References
Anderson, O.R., 1983. The radiolarian symbiosis. In: Goff, L.J. (Ed.), Algal Symbiosis: A Continuum of Interaction Strategies. Cambridge University Press, Cambridge, UK, pp. 6990. Baba, M., Takeshige, K., Baba, N., Ohsumi, Y., 1994. Ultrastructural analysis of the autophagic process in yeast, detection of autophagosomes and their characterization. J. Cell Biol. 124, 903913. Baird, A.H., Bhagooli, R., Ralph, P.J., Takahashi, S., 2009. Coral bleaching: the role of the host. Trends Ecol. Evol. 24, 1620. Baker, A.C., Coffroth, M.A., Willis, B.L., 2009. Bleaching resistance and the role of algal endosymbionts. In: van Oppen, M.J.H., Lough, J.M. (Eds.), Coral Bleaching. Patterns, Processes, Causes and Consequences. Ecological Studies Series, vol. 2005. Springer, Berlin, pp. 83101. Bernhard, J.M., 2003. Potential symbionts in bathyal foraminifera. Science 299, 861. Bishop, D.G., Bain, J.M., Downton, W.J.S., 1976. Ultrastructure and lipid composition of zooxanthellae from Tridacna maxima. Aust. J. Plant Physiol. 3, 3340. Blanco, A.V., Chapman, G.B., 1987. Ultrastructural features of the marine dinoagellate Amphidinium klebsii (Dinophyceae). Trans. Am. Microsc. Soc. 106, 201213. Bosch, T.C.G., 2008. Stem cells in immortal Hydra. In: Bosch, T.C.G. (Ed.), Stem Cells: From Hydra to Man. Springer, The Netherlands, pp. 3757. Brehelin, C., Kessler, F., van Wijk, K.J., 2007. Plastoglobules, versatile lipoprotein particles in plastids. Trends Plant Sci. 12, 260266. Brown, B.E., Le Tissier, M.D.A., Bythell, J.C., 1995. Mechanisms of bleaching deduced from histological studies of reef corals sampled during a natural bleaching event. Mar. Biol. 122, 655663. Buvat, R., 1971. Origin and continuity of cell vacuoles. In: Reinert, J., Ursprung, J.H. (Eds.), Results and Problems in Cell Differentiation, vol. 2. Springer, Berlin, pp. 127157. Buvat, R., Robert, G., 1979. Vacuole formation in the actively growing root meristem of barley (Hordeum sativum). Am. J. Bot. 66, 12191237. Carpenter, E.J., Foster, R.A., 2002. Marine cyanobacterial symbioses. In: Rai, A.N., Bergman, B., Rasmussen, U. (Eds.), Cyanobacteria in Symbiosis. Kluwer, Dordrecht, The Netherlands, pp. 1118. Cavalier-Smith, T., 1999. Principles of protein and lipid targeting in secondary symbiogenesis: euglenoid, dinoagellate, and sporozoan plastid origins and the eukaryote family tree. J. Eukaryot. Microbiol. 46, 347366. Cavalier-Smith, T., 2000. Membrane heredity and early chloroplast evolution. Trends Plant Sci. 5, 174182. Cavalier-Smith, T., 2002. Nucleomorphs: enslaved algal nuclei. Curr. Opin. Microbiol. 5, 612619. Cheung, A.Y., McNellis, T., Piekos, B., 1993. Maintenance of chloroplast components during chromoplast differentiation in the tomato mutant green esh. Plant Physiol. 101, 12231229. Chiba, A., Ishida, H., Nishizawa, N.K., Makino, A., Mae, T., 2003. Exclusion of ribulose1,5-bisphosphate carboxylase/oxygenase from chloroplasts by specic bodies in naturally senescing leaves of wheat. Plant Cell Physiol. 44, 914921. Clark, K.B., Jensen, K.R., Stirts, H.M., Fermin, C., 1981. Chloroplast symbiosis in a nonelysiid mollusc, Costasiella liljanae Marcus (Hermaeidae, Ascoglossa (=Sacoglossa)): effects of temperature, light intensity, and starvation on carbon xation rate. Biol. Bull. 160, 4354. Contento, A.L., Xiong, Y., Bassham, D.C., 2005. Visualization of autophagy in Arabidopsis using the uorescent dye monodansylcadaverine and a GFP-AtATG8e fusion protein. Plant J. 42, 598608. Crawford, M.J., Thomsen-Zieger, N., Ray, M., Schachtner, J., Roos, D.S., Seeber, F., 2006. Toxoplasma gondii scavenges host-derived lipoic acid despite its de novo synthesis in the apicoplast. EMBO J. 25, 32143222. Dahl, E.L., Rosenthal, P.J., 2007. Multiple antibiotics exert delayed effects against the Plasmodium falciparum apicoplast. Antimicrob. Agents Chemother. 51, 3485 3490. Dahl, E.L., Rosenthal, P.J., 2008. Apicoplast translation, transcription and genome replication, targets for antimalarial antibiotics. Trends Parasitol. 24, 279284.

W.G. van Doorn, K. Yoshimoto / Ageing Research Reviews 9 (2010) 117130 DellaPenna, D., Pogson, B.J., 2006. Vitamin synthesis in plants: tocopherols and carotenoids. Ann. Rev. Plant Biol. 57, 711738. Dubey, J.P., Lindsay, D.S., Speer, C.A., 1998. Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clin. Microbiol. Rev. 11, 267299. Fink, A.L., 1999. Chaperone-mediated protein folding. Physiol. Rev. 79, 425449. Fishman, Y., Zlotkin, E., Sher, D., 2008. Expulsion of symbiotic algae during feeding by the green Hydra a mechanism for regulating symbiont density? PLoS ONE 3 (7), e2603, doi:10.1371/journal.pone.0002603. Fitt, W.K., Trench, R.K., 1981. Spawning, development, and acquisition of zooxanthellae by Tridacna squamosa (Mollusca, Bivalvia). Biol. Bull. 161, 213235. Fukuda, H., Watanabe, Y., Kuriyama, H., Aoyagi, S., Sugiyama, M., Yamamoto, R., Demura, T., Minami, A., 1998. Programming of cell death during xylogenesis. J. Plant Res. 111, 253256. Gast, R.J., Moran, D.M., Dennett, M.R., Caron, D.A., 2007. Kleptoplasty in an Antarctic dinoagellate: caught in evolutionary transition? Environ. Microbiol. 9, 3945. Gimenez Casalduero, F., Muniain, C., 2008. The role of kleptoplasts in the survival rates of Elysia timida (Risso, 1818): (Sacoglossa: Opisthobranchia) during periods of food shortage. J. Exp. Mar. Biol. Ecol. 357, 181187. Goodman, C.D., Su, V., McFadden, G.I., 2007. The effects of anti-bacterials on the malaria parasite Plasmodium falciparum. Mol. Biochem. Parasitol. 152, 181191. Gray, M.W., Burger, G., Lang, B.F., 2001. The origin and early evolution of mitochondria. Genome Biol. 2 (6) review 1018.11018.5. Gunther, S., Storm, J., Muller, S., 2009. Plasmodium falciparum, organelle-specic acquisition of lipoic acid. Int. J. Biochem. Cell Biol. 41, 748752. Habetha, M., Anton-Erxleben, F., Neumann, K., Bosch, T.C.G., 2003. The Hydra viridis/ Chlorella symbiosis. Growth and sexual differentiation in polyps without symbionts. Zoology 106, 101108. Hanson, M.R., Sattarzadeh, A., 2008. Dynamic morphology of plastids and stromules in angiosperm plants. Plant Cell Environ. 31, 646657. Hirose, E., Ohtsuka, K., Ishikura, M., Maruyama, T., 2004. Ultraviolet absorption in ascidian tunic and ascidian-Prochloron symbiosis. J. Mar. Biol. Assoc. U.K. 84, 789794. Honegger, R., 1991. Functional aspects of the lichen symbiosis. Ann. Rev. Plant Biol. 42, 553578. Hopkins, J., Fowler, R., Krishna, S., Wilson, I., Mitchell, G., Bannister, L., 1999. The plastid in Plasmodium falciparum asexual blood stages: a three-dimensional ultrastructural analysis. Protist 150, 283295. Horner, H.T., Healy, R.A., Ren, G., Fritz, D., Klyne, A., Seames, C., Thornburg, R.W., 2007. Amyloplast to chromoplast conversion in developing ornamental tobacco oral nectaries provides sugar for nectar and antioxidants for protection. Am. J. Bot. 94, 1224. Hortensteiner, S., Lee, D.W., 2007. Chlorophyll catabolism and leaf coloration. In: Gan, S. (Ed.), Senescence Processes in Plants. Blackwell, Oxford, pp. 108144. Inada, N., Sakai, A., Kuroiwa, H., Kuroiwa, T., 1998. Three-dimensional analysis of the senescence program in rice (Oryza sativa L.) coleoptiles. Planta 206, 585 597. Ishida, H., Yoshimoto, K., Izumi, M., Reisen, D., Yano, Y., Makino, A., Ohsumi, Y., Hanson, M.R., Mae, T., 2008. Mobilization of Rubisco and stroma-localized uorescent proteins of chloroplasts to the vacuole by an ATG gene-dependent autophagic process. Plant Physiol. 148, 142155. Jantzen, C., Wild, C., El-Zibdah, M., Roa-Quiaoit, H.A., Haacke, C., Richter, C., 2008. Photosynthetic performance of giant clams, Tridacna maxima and T. squamosa. Red Sea Mar. Biol. 5, 211221. Johnson, M.D., Oldach, D., Delwiche, C.F., Stoecker, D.K., 2007. Retention of transcriptionally active cryptophyte nuclei by the ciliate Myrionecta rubra. Nature 445, 426428. Jones, R.J., Yellowlees, D., 1997. Regulation and control of intracellular algae (=zooxanthellae) in hard corals. Phil. Trans. R. Soc. Lond. B (Biol. Sci.) 352, 457468. Julliard, J., Douce, R., 1991. Biosynthesis of the thiazole moiety of thiamin (Vitamin B1) in higher plant chloroplasts. Proc. Natl. Acad. Sci. U.S.A. 88, 20422045. Keeling, P.J., 2004. Diversity and evolutionary history of plastids and their hosts. Am. J. Bot. 91, 14811493. Kempf, S.C., 1984. Symbiosis between the zooxanthella Symbiodinium(=Gymnodinium) microadriaticum (Freudenthal) and four species of nudibranch. Biol. Bull. 166, 110126. Kikinger, R., 2008. Cotylorhiza tuberculata (Cnidaria: Scyphozoa) life history of a stationary population. Mar. Ecol. 13, 333362. Kim, I., Rodriguez-Enriquez, S., Lemasters, J.J., 2007. Selective degradation of mitochondria by mitophagy. Arch. Biochem. Biophys. 462, 245253. Krupinska, K., 2007. Fate and activities of plastids during leaf senescence. In: Wise, R.R., Hoober, J.K. (Eds.), The Structure and Function of Plastids. Springer, Dordrecht, pp. 433449. Kuriyama, H., 1999. Loss of tonoplast integrity programmed in tracheary element differentiation. Plant Physiol. 121, 763774. Kvennefors, E.C.E., Roff, G., 2009. Evidence of cyanobacteria-like endosymbionts in Acroporid corals from the Great Barrier Reef. Coral Reefs 28, 547. ` ` Ladriere, O., Compere, P., Decloux, N., Vandewalle, P., Poulicek, M., 2008. Morphological alterations of zooxanthellae in bleached cnidarian hosts. Cah. Biol. Mar. 49, 215227. Leon, P., Arroyo, A., Mackenzie, S., 1998. Nuclear control of plastid and mitochondrial development in higher plants. Ann. Rev. Plant Physiol. Plant Mol. Biol. 49, 453480. Lesser, M.P., Shick, J.M., 1990. Effects of visible and UV radiation on the ultrastructure of zooxanthellae Symbiodinium sp. in culture and in situ. Cell Tissue Res. 261, 501508.

129

Lesser, M.P., Mazel, C.H., Gorbunov, M.Y., Falkowski, P.G., 2004. Discovery of nitrogen-xing cyanobacteria in corals. Science 305, 9971000. Lim, P.O., Kim, H.J., Nam, H.G., 2007. Leaf senescence. Ann. Rev. Plant Biol. 58, 115 136. Liu, Y., Schiff, M., Czymmek, K., Talloczy, Z., Levine, B., Dinesh-Kumar, S.P., 2005. Autophagy regulates programmed cell death during the plant innate immune response. Cell 121, 567577. Ljubesic, N., Wroscher, M., Devide, Z., 1991. Chromoplasts the last stages in plastid development. Int. J. Dev. Biol. 35, 251258. Lough, J.M., van Oppen, M.J.H., 2009. Introduction: coral bleaching patterns, processes, causes and consequences. In: van Oppen, M.J.H., Lough, J.M. (Eds.), Coral Bleaching. Patterns, Processes, Causes and Consequences. Ecological Studies Series, vol. 2005. Springer, Berlin, pp. 14. Ludwig, L.D., 1969. Die Zooxanthellen bei Cassiopeia andromeda Eschscholz 1929 (Polyp-stadium) und ihre Bedeutung fur die Strobilation. Zool. Jahrb. Abt. Anat. 86, 238277. Marshall, P., Schuttenberg, H., 2006. A Reef Managers Guide to Coral Bleaching. Australian Government. Great Barrier Reef Marine Park Authority, Townsville, Australia. Martn, M., Casano, L.M., Zapata, J.M., Guera, A., del Campo, E.M., Schmitz-Linneweber, C., Maier, R.M., Sabater, B., 2004. Role of thylakoid Ndh complex and peroxidase in the protection against photo-oxidative stress: uorescence and enzyme activities in wild-type and ndhF-decient tobacco. Physiol. Plant. 122, 443452. Martnez, D.E., 1997. Mortality patterns suggest lack of senescence in hydra. Exp. Gerontol. 33, 217225. Martnez, D.E., Costa, M.L., Gomez, F.M., Otegui, M.S., Guiamet, J.J., 2008. Senescence-associated vacuoles are involved in the degradation of chloroplast proteins in tobacco leaves. Plant J. 56, 196206. Marty, F., 1978. Cytochemical studies on GERL, provacuoles, and vacuoles in root meristematic cells of Euphorbia. Proc. Natl. Acad. Sci. U.S.A. 75, 852856. Marty, F., 1999. Plant vacuoles. Plant Cell 11, 587599. Matile, P., 2001. Senescence and cell death in plant development: chloroplast senescence and its regulation. In: Aro, E.M., Andersson, B. (Eds.), Regulation of Photosynthesis. Kluwer, Dordrecht, Netherlands, pp. 277296. Mazumdar, J., Wilson, H., Mazek, K., Junter, C.A., Striepen, B., 2006. Apicoplast fatty acid synthesis is essential for organelle biogenesis and parasite survival in Toxoplasma gondii. Proc. Natl. Acad. Sci. U.S.A. 103, 1319213197. McCoy, A.M., Balzer, I., 2004. Algal symbiosis in atworms. In: Seckbach, J. (Ed.), Symbiosis. Springer, The Netherlands, pp. 559574. Michaels, A.F., Silver, M.W., 1988. Primary production, sinking uxes and the microbial food web. Deep Sea Res. 35, 473490. Minamikawa, T., Toyooka, K., Okamoto, T., Hara-Nishimura, I., Nishimura, M., 2001. Degradation of ribulose-bisphosphate carboxylase by vacuolar enzymes of senescing French bean leaves: immunocytochemical and ultrastructural observations. Protoplasma 218, 144153. Minnhagen, S., Carvalho, W.F., Salomon, P.S., Janson, S., 2008. Chloroplast DNA content in Dinophysis (Dinophyceae) from different cell cycle stages is consistent with kleptoplasty. Environ. Microbiol. 10, 24112417. Mizushima, N., 2005. The pleiotropic role of autophagy: from protein metabolism to bactericide. Cell Death Differ. 12, 15351541. Mizushima, N., 2007. Autophagy: process and function. Genes Dev. 21, 28612873. Mendes Lopes, R., Silveira, M., 2004. Symbiosis between a pelagic atworm and a dinoagellate from a tropical area: structural observations. Hydrobiologia 287, 277284. Menzies, R.A., Gold, P.H., 1971. The turnover of mitochondria in a variety of tissues of young adult and aged rats. J. Biol. Chem. 246, 24252429. Mondy, W.L., Pierce, S.K., 2003. Apoptotic-like morphology is associated with annual synchronized death in kleptoplastic sea slugs (Elysia chlorotica). Invertebrate Biol. 122, 126137. Moore, R.B., Obornk, M., Janouskovec, J., Chrudimsky, T., Vancova, M., Green, D.H., Wright, S.W., Davies, N.W., Bolch, C.J., Heimann, K., Slapeta, J., Hoegh-Guldberg, O., Logsdon, J.M., Carter, D.A., 2008. A photosynthetic alveolate closely related to apicomplexan parasites. Nature 451, 959963; 452, 900. Mujer, C.V., Andrews, D.L., Manhart, J.R., Pierce, S.K., Rumpho, M.E., 1996. Chloroplast genes are expressed during intracellular symbiotic association of Vaucheria litorea plastids with the sea slug Elysia chlorotica. Cell Biol. 93, 12333 12338. Muller-Parke, G., Lee, K.W., Cook, C.B., 2008. Changes in the ultrastrucure of symbiotic zooxhanthellae (Symbiodinium sp., Dinophyceae) in fed and starced sea anemones maintained under high and low light. J. Phycol. 32, 987 994. Munne-Bosch, S., 2007. Aging in perennials. Crit. Rev. Plant Sci. 26, 123138. Muscatine, L., Lenhoff, H.M., 1965. Symbiosis of hydra and algae. II. Effects of limited food and starvation on growth of symbiotic and aposymbiotic hydra. Biol. Bull. 129, 316328. Natesan, S.K.A., Sullivan, J.A., Gray, J.A.C., 2005. Stromules: a characteristic cellspecic feature of plastid morphology. J. Exp. Bot. 56, 787797. Nooden, L.D., Penney, J.P., 2001. Correlative controls of senescence and plant death in Arabidopsis thaliana (Brassicaceae). J. Exp. Bot. 52, 21512159. Obornk, M., Janouskovec, J., Chrudimsky, Y., Lukes, J., 2009. Evolution of the apicoplast and its hosts: from heterotrophy to autotrophy and back again. Int. J. Parasitol. 39, 112. Otegui, M.S., Noh, Y.S., Martnez, D.E., Vila Petroff, M.G., Staehelin, L.A., Amasino, R.N., Guiamet, J.J., 2005. Senescence-associated vacuoles with intense proteolytic activity develop in leaves of Arabidopsis and soybean. Plant J. 41, 831844.

130

W.G. van Doorn, K. Yoshimoto / Ageing Research Reviews 9 (2010) 117130 Taylor, D.L., 1978. Articially induced symbiosis between marine agellates and vertebrate tissues in culture. J. Eukar. Microbiol. 25, 7781. Tchernov, D., Gorbunov, M.Y., de Vargas, C., Yadav, S.N., Milligan, A.J., Haggblom, M., Falkowski, P.G., 2004. Membrane lipids of symbiotic algae are diagnostic of sensitivity to thermal bleaching in corals. Proc. Natl. Acad. Sci. U.S.A. 101, 1353113535. Thompson, A.R., Doelling, J.H., Suttangkakul, A., Vierstra, R.D., 2005. Autophagic nutrient recycling in Arabidopsis directed by the ATG8 and ATG12 conjugation pathways. Plant Physiol. 138, 20972110. Trench, R.K., 1979. The cell biology of plantanimal symbiosis. Ann. Rev. Plant Physiol. 30, 485531. Trench, R.K., Boyle, J.E., Smith, D.C., 1973. The association between chloroplasts of Codium fragile and the mollusc Elysia viridis. II. Chloroplast ultrastructure and photosynthetic carbon xation in E. viridis. Proc. R. Soc. Lond. B 184, 6381. van Doorn, W.G., 2006. Plant programmed cell death and the point of no return. Trends Plant Sci. 10, 478483. van Doorn, W.G., Woltering, E.J., 2005. Many ways to exit? Cell death categories in plants. Trends Plant Sci. 10, 117122. Venn, A.A., Loram, J.E., Douglas, A.E., 2008. Photosynthetic symbioses in animals. J. Exp. Bot. 59, 10691080. Wada, S., Ishida, H., Izumi, M., Yoshimoto, K., Ohsumi, Y., Mae, T., Makino, A., 2009. Autophagy plays a role in chloroplast degradation during senescence in individually darkened leaves. Plant Physiol. 49, 885893. Wakeeld, T.S., Farmer, M.A., Kempf, S.C., 2000. Revised description of the ne structure of in situ zooxanthellae genus Symbiodinium. Biol. Bull. 199, 7684. Waller, R.F., McFadden, G.I., 2005. The apicoplast: a review of the derived plastid of apicomplexan parasites. Curr. Issues Mol. Biol. 7, 5779. Waller, R.F., Reed, M.B., Cowman, A.F., McFadden, G.I., 2000. Protein trafcking to the plastid of Plasmodium falciparum is via the secretory pathway. EMBO J. 19, 17941802. Wise, R.R., 2007. The diversity of plastid form and function. In: Wise, R.R., Hoober, R.K. (Eds.), Advances in Photosynthesis and Respiration, Volume 23. The Structure and Function of Plastids. Springer, Amsterdam, pp. 226. Wittenbach, V.A., Lin, W., Hebert, R.R., 1982. Vacuolar localization of proteases and degradation of chloroplasts in mesophyll protoplasts from senescing primary wheat leaves. Plant Physiol. 69, 98102. Xie, Z., Klionsky, D.J., 2007. Autophagosome formation: core machinery and adaptations. Nat. Cell Biol. 9, 11021109. Yakovleva, I.M., Baird, A.H., Yamamoto, H.H., Bhagooli, R., Nonaka, M., Hidaka, M., 2009. Algal symbionts increase oxidative damage and death in coral larvae at high temperature. Mar. Ecol. Prog. Ser. 378, 105112. Yoshimoto, K., Hanaoka, H., Sato, S., Kato, T., Tabata, S., Noda, T., Ohsumi, Y., 2004. Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy. Plant Cell 16, 29672983. Zapata, J.M., Guera, A., Esteban-Carrasco, A., Martn, M., Sabater, B., 2005. Chloroplasts regulate leaf senescence: delayed senescence in transgenic ndhF-defective tobacco. Cell Death Diff. 12, 12771284.

Park, M.G., Park, J.S., Kim, M., Yih, W., 2008. Plastid dynamics during survival of Dinophysis caudata without its ciliate prey. J. Phycol. 44, 11541163. Peoples, M.B., Beilharz, V.C., Waters, S.P., Simpson, R.J., Dalling, M.J., 1980. Nitrogen redistribution during grain growth in wheat (Triticum aestivum L.). Chloroplast senescence and the degradation of ribulose-1,5-bisphosphate carboxylase. Planta 149, 241251. Phillips, M.A., Leon, P., Boronat, A., Rodriguez-Concepcion, M., 2008. The plastidial MEP pathway: unied nomenclature and resources. Trends Plant Sci. 13, 619 623. Pratchett, M.S., Wilson, S.K., Graham, N.A.J., Munday, P.L., Jones, G.P., Polunin, N.V.C., 2009. Coral bleaching and consequences for motile reef organisms: past, present and uncertain future effects. In: van Oppen, M.J.H., Lough, J.M. (Eds.), Coral Bleaching. Patterns, Processes, Causes and Consequences. Ecological Studies Series, vol. 2005. Springer, Berlin, pp. 139157. Provasoli, L., Yamasu, T., Manton, I., 1968. Experiments on the resynthesis of symbiosis in Convoluta roscoffensis with different agellate cultures. J. Mar. Biol. Assoc. U.K. 48, 465479. Pyke, K., 2009. Plastid Biology. Cambridge University Press, Cambridge, UK. Ramya, T.N.C., Mishra, S., Karmodiya, K., Surolia, N., Surolia, A., 2007. Inhibitors of nonhousekeeping functions of the apicoplast defy delayed death in Plasmodium falciparum. Antimicrob. Agents Chemother. 51, 307316. Reyes-Prieto, A., Weber, A.P.M., Bhattacharya, D., 2007. The origin and establishment of the plastid in algae and plants. Ann. Rev. Genet. 41, 147168. Rumpho, M.E., Dastoor, F.P., Manhart, J.R., Lee, J., 2006. The kleptoplast. In: Wise, R.R., Hoober, J.K. (Eds.), The Structure and Function of Plastids. Springer, Dordrecht, pp. 451473. Rumpho, M.E., Worful, J.M., Lee, J., Kannan, K., Tyler, M.S., Bhattacharya, D., Moustafa, A., Manhart, J.R., 2008. Horizontal gene transfer of the algal nuclear gene psbO to the photosynthetic sea slug Elysia chlorotica. Proc. Natl. Acad. Sci. U.S.A. 105, 1758917590. Sakamoto, W., 2006. Protein degradation machineries in plastids. Ann. Rev. Plant Biol. 57, 599621. Schultz, G., Soll, J., Fiedler, E., Schulze-Siebert, D., 1985. Synthesis of prenylquinones in chloroplasts. Physiol. Plant. 64, 123129. Seo, S., Okamoto, M., Iwai, T., Iwano, M., Fukui, K., Isogai, A., Nakajima, N., Ohashi, N., 2000. Reduced levels of chloroplast FtsH protein in tobacco mosaic virusinfected tobacco leaves accelerate the hypersensitive response. Plant Cell 12, 917932. Stoecker, D.K., Silver, M.W., 1990. Replacement and aging of chloroplasts in Strombidium capitatum (Ciliophora, Oligotrichida). Mar. Biol. 107, 491502. Stoecker, D.K., Swanberg, N., Tyler, S., 1989. Oceanic mixotrophic atworms. Mar. Ecol. Prog. Ser. 58, 4151. Sugiura, Y., 1964. On the life-history of rhizostome medusae. II. Indispensability of zooxanthellae for strobilation in Mastigias papua. Embryologia 8, 223233. Taylor, D.L., 1971. On the symbiosis between Amphidinium klebsii (Dinophyceae) and Atnphiscolops langerhansi (Turbellaria: Acoela). J. Mar. Biol. Assoc. U.K. 51, 301313.