LEC 3.

ASCENT OF SAP
B.Sc BIOTECHNOLOGY 08.05.2012 TRANSLOCATION OF WATER

Introduction
Plants absorb water through the roots. This absorbed water reached up to leaves through the stem branches. Most of the water is lost by the transpiration from the leaves. Only small amount of water utilized in vital activities of the plant.

 "The upward movement of absorbed water by the under ground roots towards aerial parts of the plant against the gravitation is called Ascent of sap". Process of ascent of sap takes place from a small plant to a quite large plant e.g. in Sequoia semipervense. It has been experimentally proved that water moves upwards through the vessels and tracheids of xylem.

Experiment No. 1 - Balsam

Take aqueous solution of eosine in a beaker and immerse the cut end of (stock) of balsam plant in it. After sometimes strips of red colour are seen in the stem. The transverse section of this stem confirms because only vessels and tracheids are stained with eosine dye. So that it is proved that the ascent of sap take place through the xylem

Experiment No. 2 - Ringing Experiment

First of all ringing experiment was carried by Hartig, Malpighi and Stephen Hales. We take a branch with leaves immersed in beaker which is filled with water. Now remove all the tissues which are present above the xylem [cortex, cambium and phloem]. Now placed this experiment in a suitable environment for 2 - 3 days. Then we observe that leaves did not wilt. This experiment makes it clear that water moves up only through vessels and tracheids of the xylem.

 Ringing [girdling] experiment has been done only in dicotyledons because their vascular bundles lies in a ring. While vascular bundles are scattered in monocotyledons, so that this experiment can not be possible.

Structures involved in Ascent of Sap: Various experiments like girdling, staining and plugging, indicate that the xylem tissue is mainly responsible for the movement of water. As xylem consists of tracheae otherwise called vessels, they form a system of fine channels running from roots to all other regions of the plant body and form a beautifully branched supply system, which is almost similar to that of arteries in animals.

Theories of Ascent of sap

Many theories have been put forwards by various co-workers to explain the mechanism of ascent of sap. (1) Vital Force Theory (2) Root Pressure Theory (3) Physical Force Theory

(1) Vital force theory

Vital theories presumes that ascent of sap is due to the vital activity of living cells of the plants. The opinion of the various scientist for ascent of sap as follows :

(A) Westermaier's theory

Westermaier was the first man who proposed his view about the vital force theory. He explained that force for upward conduction of water is provided by the xylem parenchyma cells and vessels and tracheids are simply acting as water reservoirs.

(B) Theory of Godlewski

He proposed that ascent of sap is brought about by periodic changes in osmotic pressure of xylem parenchyma and medullary rays. A pumping action starts as a result of this changes that causes ascent of sap. So that a sort of stair case movement of water took place in the plant. Thus it is known as "Relay pump theory" or "Clambering theory". Osmotic pressure of parenchyma increased, water from the xylem vessel diffused into it. this water moves up because of atmospheric pressure during this period, value of osmotic pressure of parenchyma decreased (lower down) and hence this water recieved by xylem which is lies at higher level.

(C) Pulsation theory of J.C. Bose

J.C. Bose is known as father of Indian plant physiology. He proposed pulsation theory. He assumed that the pulsatory activity like heart beat is found in the inner cortical cells which are situated just above the endodermis. Resulting of this pulsatory activity, water forced or pumped into cavities (Vessel) of xylem and water moves upward. He conducted his experiment on a plant Desmodium gyrans (Indian telegraph plant) which is member of Papillionaceae family.

 J.C. Bose measured this pulsation activity by "Electric probe" (Galvanometer) apparatus Molish A scientist named Molish who supported the pulsation theory of J.C. Bose and he introduced a detail description of pulsation theory. According to him the pulsatory activity increased to 14 seconds by the application of narcotics drugs on the plant.

Objections
The view of vital theory discarded by Straburger and Boucheries. They proved by their experiment that the living cells are not essential for ascent of sap. Because when the living cells are killed (destroyed) by picric acid or HgCl2 solution even then ascent of sap continues.

(2) Root Pressure Theory

This theory was proposed by Priestley. Root pressure : A positive pressure is present in the xylem sap of roots. The term root pressure was coined by Stephan Hales. Priestley called it "hydrostatic pressure" Root pressure is also known as 'exudation pressure'. The highest value of root pressure is found in those plants which are growing in well aerated and sufficient amount of soil during the humidity in the atmosphere. Root pressure usually develops during the night when absorption is maximum and transpiration is minimum.

 The maximum value of root pressure falls in the range of 2 - 3 atmosphere. The liquid which flows from the freshly decapitated stem of the plant is called bleeding. This is also due to root pressure. Guttation is also takes place due to root pressure. Ascent of sap through the root pressures only possible in small and herbaceous plants. Therefore it is only applicable for small and herbaceous plants.

 The process of ascent of sap can not be explained by root pressure because it has following limitations (1) Gymnosperms lack root pressure and the conifers are very tall plants. (2) The maximum transpiration is found during the summer and plants require more water but in that time root pressure is found to be very less. (3) The rate of ascent of sap is found rapidly even in the absence of root pressure. (4) The rising up of water can not be possible at 2 atmosphere in tall plants. Upto 200 feet height plants require 10 atmosphere root pressure to push the water to the top of the plants. (5) A negative root pressure is found in some of the desert plants. (6) Root pressure is found less in summer and more in winter. (7) Root pressure takes place in the special conditions when the rate of absorption is higher and transpiration rate is minimum. Hence, root pressure is not significant in most of the plants. It is only effective in low transpiring herbaceous plants. Therefore, theory explain partially the mechanism of ascent of sap.

(3) Physical Force Theory

According to this theory, the ascent of sap takes place by the vessels and tracheids through some physical forces. The following theories stated under this heading. (A) Capillary force theory : This theory was proposed by Boehm. According to him the vessels and tracheid which are present in xylem, acts as capillaries and water rises up by surface tension in their capillaries.

 Limitation of this theory as follows : (a) Xylem vessels are not freely immersed into the water. (b) The ends of vessels and tracheids are closed while ends of capillaries are open. (c) The diameter of the capillaries should be very thin because capillary with narrow lumen encourages the entry of more water. But in tall plants the diameter of vessels and tracheids is more. At this height capillary force does not operate. (d) Tall plants like Gymnosperms having tracheids instead of vessels which have many transverse septum. Therefore, tracheids are homologous as capillary. But still ascent of sap occurs. (e) Lifting power of capillary is not much and can not account for the rise of water exceeding 3 meters. It is capable for few centimeters rise only. Capillary theory is completely discarded on the basis of above reasons.

(B) Imbibition force theory

This theory was proposed by Von Sachs. According to him water rises up in the wall of the xylem cells through the imbibition end through their lumen. But now it has been clear that water rises up only through the lumen not by the walls. The wall of vessels and tracheids of xylem are lignified and lignin is impermeable to water. Thus this theory also discarded.

(C) Chain theory

This theory proposed by Jamin. According to this theory, the molecules of water and air are arranged alternatively to form a chain. Ascent of sap takes place due to spreading of air in the chain.

(D) Transpiration pull - cohesion force theory

This theory also known as "Water column theory" or "Cohesive force theory" This theory was proposed by Dixon and Jolly. This is the most accepted theory at the present time and it accounts satisfactory explanation for the rising of water. This theory is based on the following facts :

(A) Transpiration pull

Water is lost continuously from the leaves of the plants as a result of transpiration. Water vapour move out from the leaves. As a result of loss of water from the mesophyll cell. Diffusion pressure deficit always remain higher in the mesophyll cells. Due to this mesophyll cells absorb water from adjoining internal mesophyll cells and compensate this loss of water. This loss ultimately compensating by xylem which leads water deficit in the xylem. Rapid transpiration develops a pull or tension in xylem which is called transpiration pull. It is about 20 atmospheres. This pull is called "Negative pressure" because it is develops from aerial parts to the under ground part of the plants.

(b) Cohesion Force of Water

A force of mutual attraction present between the water molecules is called "Cohesive force". Water molecules are held together continuously by cohesive force and to form a continuous water column. This cohesive force is up to 45 . 270 atmosphere.

(c) Adhesive Force

Water molecules are also attached with the wall of vessels and tracheids through a force called adhesive force. Both forces work together and maintain the continuity in between water and cell wall. Cohesive force and adhesive force works continuously in the cavity (lumen) of xylem. Both the forces are responsible for maintaining unbroken continuity of water column from the roots to the leaves. MacDougal called as "Hydrostatic system" of this continuous water column. This water column pulled upwards continuously without breaking from the roots to the leaves by transpiration. Vessels and tracheids of xylem workds as pipe (Tubes).

 The process of ascent of sap a continues through the medium of above mechanisms. Evidences in support of Dixon's theory : (1) Normally, the rate of absorption is equal to the rate of transpiration water does not come out from the cut end of the stem during transpiration. When this cut end is watered, it absorb water inside the stem. This justifies the water tension in the stem. (2) McDougal is found that daily contraction in stem of the trees with the help dendrograph apparatus. Maximum contraction is found during a maximum transpiration. (3) The maximum value of osmotic pressure of the cells of leaves is found during noon. Because the amount of water is minimum in that time

Objection to Dixon's theory

(1) The presence of air bubbles in the xylem which may break the continuity of water column. (2) Ascent of sap continues even after overlapping cuts. * Removal of objections :

 Although air bubbles are present in the xylem, but they are unable to move much distance because (i) xylem vessels do not continue for long distances, pits are present between them. (ii) Tracheid are also present along the vessels as alternative path. Pits are also present in them. (iii) Membrane and wall is present on the pits which is permeable. to water, but air bubbles do not pass out through them. Air bubbles may dissolve in water. Root pressure also helps to remove air bubbles.

 The rate of transpiration is higher than that of rate of ascent of sap during the day time and is called absorption lag. Russel and Wooley found the ratio 50 : 1 by comparative study of water movement in apoplast and symplast. The origin of root pressure is an active process. Factors affecting Ascent of sap (1) Amount of water in soil. (2) High temperature (3) High atmospheric humidity (4) High atmospheric pressure (5) Wind velocity (6) Number of stomata in leaves

Mechanism of Ascent of Sap

VITAL THEORY Suggestions have been made that living cells adjacent to conducting pipes of xylem are involved in the translocation of sap.These living cells are postulated to show some kind of pulsating action(just lioke heart beat) and pulsation of cells at successive intervals lifts the sap up. Some proponents of vital theory are 1669-Ray-Sap Circulates in Plants like blood in Animals 1884-Godlewski-Cell pump water 1927-Bose-Sap ascends by pulsation of cortical movements 1938-Dixon and Barlee-Leaves Exude water by vital action of Leaf cells

 However anatomical studies have failed to show any pumping action in the Xylem.Further numerous experiments have demonstrated clearly that living cells are not involved in the translocation of sap in the stem.

ROOT PRESSURE THEORY Acc to Root pressure theory,the positive pressure developed in the roots pumps the sap up in the xyulem ducts.This theory seems to be applcable to many species,which generate considerable amount of root pressure.For example,Grapevines have been shown to generate root pressure up to 5-6 bars,which can support a water column upto 150-160 ft

 However, root pressure cannot account for sap translocation in many situations.In many species,root pressure has not been observed at all and in many others,it is low that it cannot account for upward translocation even upto a few ft.

 COHESION THEORY BY IRISH BOTANIST HENRI DIXON and a Physicist Jon Joley(1894) Supported by E.Askenasy(1895)and Renner(1895).

 Water has a high Cohesive( Mutual attraction) force.This is primarily because of the formation of Hydrogen bonds between water molecules.Water also shows forces of adhesive with the walls of xylem vessels and tracheids.Because of Cohesive and adhesive forces,it exists as a column in the xylem elements.

Organismal Circulation
Unicellular Organisms Autotrophic Multicellular Organisms (Heterotrophic Multicellular Organisms)

Cyclosis in Physarum polycephalum, a slime mold

This organism consists of one very large cytoplasm (plasmodium) with many nuclei and food vacuoles in the cytosol (coenocytic). Slime molds can weigh up toward kilogram range and moves their blob-like mass around exclusively by cyclosis. Here you can see, in a thin region of cytoplasm, that it moves along pathways that are river-like in appearance.

Transport is NOT always unidirectional.

http://botit.botany.wisc.edu/courses/img/Botany_1 30/Movies/Slime_mold.mov

The correct taxonomic affiliation is unclear. It has been treated as Fungus and Protist. Further study is needed to resolve its position.

What is the ATP source?

Cyclosis: cytoplasmic streamingintracellular circulation

Chloroplasts and other organelles have surface proteins with myosinlike activity. Elodea canadensis ATP and Calcium allow myosin to slide along actin filaments, resulting in circulation of organelles within the cell.

Microfilaments of actin are found just under cell membrane.

QuickTime an d a Cinepak decompressor are need ed to see this p icture .

http://www.microscopy-uk.org.uk/mag/imgnov00/cycloa3i.avi

What is the source of ATP?

Can you be more specific?

If light intensity were reduced, what would be the prediction on rate of cyclosis based on your hypothesis?

Figure 36-3 Page 793

The shoot organ system is photoautotrophic, taking in CO2 and releasing O2 in daylight.
Diffusion is sufficient to exchange gases. But solutes need to be circulated in the large plant body as diffusion is too slow!!

Node Internode Node Leaves

Apical bud Axillary bud CO2 in and O2 out

Branch O2 in and CO2 out

Stem

Lateral roots

The root organ system is chemoheterotrophic, taking in O2 and releasing CO2 in the darkness of the soil environment.

O2 in and CO2 out Taproot

Figure 36-3 Page 793

The shoot system produces carbohydrates (etc.) by photosynthesis. These solutes are transported to the roots in the phloem tissue:

Node Internode Node Leaves

Apical bud Axillary bud Carbohydrate etc.

Branch

Translocation
Stem Transpiration
The root system removes water and minerals from the soil environment. These solutes are transported to the shoot in the xylem tissue:

Translocation

Lateral roots

Transpiration

Water and Minerals

Taproot

Figure 36-3 Page 793

Node Internode

Apical bud Axillary bud Carbohydrate etc.

Because these pathways involve solutes in water passing in the adjacent tissues of a narrow vascular bundle, this is a circulation system!

Node Leaves Branch

Transpiration and Translocation The water is moving up the xylem, and down the phloem, making a full circuit!

Stem Transpiration

Translocation

Lateral roots

Water and Minerals

Taproot

Figure 36-18 Page 802

Plants occur in two major groups (and some minor ones)

They differ, in part, in their circulation systems:

Cross section of a eudicot stem

Cross section of a monocot stem

Epidermis

Cortex Pith

Ground tissue

Vascular bundles

Dicots initially have one ring of vascular bundles

Monocots rapidly develop multiple, concentric, rings of vascular bundles

Monocot stem anatomy

Mature Monocot
Young Monocot

vascular bundles

As a monocot plant grows in diameter, new bundles are added toward the outside for increased circulation to the larger plant body.

Monocot stem anatomy

Is this slice from a young or a mature part of the corn stem? Lets take a closer look at the vascular tissues

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Monocot stem anatomy: vascular bundle

Translocation

Transpiration

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Why must xylem do a lot more transport than phloem?

Dicot circulation: stem anatomy

Dicots start with one ring of bundles

Lets take a closer look at the vascular tissues

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Dicot stem anatomy: vascular bundle

phloem fibers
Support of Stem functional phloem Translocation vascular cambium Cell Divison: More Xylem and Phloem

xylem

Transpiration

As a dicot grows, how does it add vascular capacity to become a tree?

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Dicot stem anatomy: vascular cambium adds secondary tissues

epidermis cortex 1 phloem 2 phloem cambium 2 xylem 1 xylem pith

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Dicot stem anatomy: vascular cambium adds secondary tissues

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Each year the vascular cambium make a new layer of secondary xylem and secondary phloem

Dicot stem anatomy: four year-old stem (3 annual growth rings) phloem etc. = bark All of these tissues were added by the vascular cambium!

xylem = wood

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Figure 36.29 Page 810 See also part (a)

or less competition in forest?

cambium phloem

or more competition in forest?

Figure 36.0 Page 791

periderm phloem cambium = bark

heartwood

pith

Two Xylem Conducting Cells: tracheid developmental sequence

Annular Helical Pitted

When flowering plants are young, water needs are limited, tracheids suffice. The walls are strengthened with secondary thickenings including lignin. Protoxylem have stretchable annular or helical thickenings.

Metaxylem have reticulate or pitted and fully rigid walls.

Tracheids have end walls and flow between cells is through pits.

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Two Xylem Conducting Cells: xylem vessel evolution plesiomorphic apomorphic

Compare Fig. 36.26 Page 806

As flowering plants age and grow, water needs increase, and tracheids need to be supplemented.
Flowering plants evolved xylem cells with larger cell diameter and perforated end walls to increase water flow. Vessels have perforated end walls or lack end walls, but lateral flow between cells is still through pits.

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Dicot stem anatomy: xylem parenchyma, vessels, and tracheids

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The huge vessel transports lots of water longitudinally, and shows lots of pits for lateral transport

Dicot stem anatomy: xylem parenchyma, vessels, and tracheids

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The huge vessel transports lots of water longitudinally, and shows lots of pits for lateral transport

Secondary xylem: cross sections of three different species

Vessels, Tracheids have different distribution patterns. Some produce big vessels only in spring wood Others produce vessels year-round.

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Dicot stem anatomy: woody stem circulation

This sketch is showing the importance of lateral transport. In both transpiration and translocation materials must move radially to the interior and to the exterior as well as up and down the plant. O2 in and CO2 out

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Dicot stem anatomy: 2-year old stem showing ray and periderm

phloem
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Rays transport sugar from the phloem toward the interior to keep pith and xylem parenchyma fueled. Rays transport water and minerals from the xylem to the exterior to keep the periderm, cortex, and phloem parenchyma hydrated.

Xylem and Phloem: tissues with many cell types but conduction function toward pith

main transpiration flow

radial transport

toward cortex

main translocation flow

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Mendocino Tree (Coastal Redwood) Sequoia sempervirens Ukiah, California

112 m tall (367.5 feet)! This tree is more than ten times taller than is theoretically possible based solely upon the length of the column of uncavitated water. How could this be achieved?

http://www.nearctica.com/trees/conifer/tsuga/Ssemp10.jpg

Transpiration in a tall tree has at least 3 critical components:

Evaporation: pulling up water from above Capillarity: climbing up of water within xylem

Root Pressure: pushing up water from below

Transpiration: root pressure (osmotic push) Solutes from translocation of sugars accumulate in roots.
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Water from the soil moves in by osmosis. Accumulating water in the root rises in the xylem. Water escapes from hydathodes.

guttation

This is not dew condensing!

Transpiration: root pressure (osmotic push) The veins (coarse and fine) show that no cell in a leaf is far from xylem and phloem (i.e.water and food!). The xylem of the veins leaks at the leaf margin in a modified stoma called the hydathode. These droplets are xylem sap.

Root pressure accounts for maybe a half-meter of push up a tree trunk.

http://img.fotocommunity.com/photos/8489473.jpg

Capillarity: maximum height of unbroken water column glass tube vacuum created gravity pulls water down

The small diameter of vessels and tracheids and the surface tension of water provide capillary (climb).
10.4m

atmospheric pressure keeps water in tube

Cohesion of water, caused by hydrogen bonds, helps avoid cavitation. A tree taller than 10.4 m would need some adaptations to avoid cavitation

water

Conifer stem anatomy: pine xylem tracheids with pits, xylem rays

vascular cambium

tracheids with pits

In spite of the limitations of tracheidsonly xylem, conifers are among the tallest of trees!

ray parenchyma

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Conifer stem anatomy: bordered pits as check-valve for flow

P low

P high

These pit features allow conifers to be very tall and still avoid cavitation in their xylem cells. As pressures change between adjacent cells, the torus movement blocks catastrophic flow that would result in cavitation.

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Transpiration: evaporation (pull)

water Water evaporating from a porous clay cap also lifts the mercury! mercury

Transpiration can lift the vacuum mercury above its normal cavitation height!

76 cm
mercury

Grown in 32PO4 (radioactive phosphorus) 1 hour

Cold medium 6 hours

Cold medium 90 hours

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new growth black

Is phosphate uptake from soil: transpiration or translocation? In xylem or phloem?

Is phosphate mobilization from lower leaf: transpiration or translocation? In xylem or phloem?

Translocation: How solutes move in phloem

Leaf

High Pressure

plasmodesmata

Root active transport

Low Pressure
Modified from: 1996 Norton Presentation Maker, W. W. Norton & Company

Translocation: How solutes move bidirectionally in phloem

Low Pressure

Developing leaves, apical bud, flowers fruits

Leaf sugars amino acids

High Pressure

Lateral buds, stems, roots, root tip Low Pressure

Modified from: 1996 Norton Presentation Maker, W. W. Norton & Company

Transpiration Evaporation: Water evaporates from mesophyll into atmosphere. Water molecules are pulled up the xylem by virtue of cohesion. Capillarity: Water climbs in the xylem cell walls by adhesion. Water molecules follow by cohesion. Root Pressure: Water moves into the root because of solutes from phloem. Pressure pushes the water up the stem. Node Internode Node Leaves

Apical bud Axillary bud Carbohydrate etc.

Branch

Stem Transpiration

Translocation

Lateral roots

Water and Minerals

Taproot
Figure 36-3 Page 793

Node Internode Node Leaves

Apical bud Axillary bud Carbohydrate etc.

Translocation

Branch

Leaf = Source Photosynthesis produces solutes. Solutes loaded into phloem by active transport. Water follows by osmosis, increasing pressure.
Root (etc.) = Sinks Solutes removed from phloem by active transport. Water follows by osmosis, reducing pressure. Pressure = Bulk Flow The pressure gradient forces phloem sap away from leaves to all sinks (bidirectionally).

Stem Transpiration

Translocation

Lateral roots

Water and Minerals

Taproot
Figure 36-3 Page 793

Thank you!
Next Lecture: TRANSPIRATION