In Vitro Cell. Dev. Biol.

30P:175-180, October 1994

1994 Society for In Vitro Biology
1054-5476/94 $02.50+0.00

Invited Review
IN VITRO SELECTION FOR SALT TOLERANCE IN CROP PLANTS:
THEORETICAL AND PRACTICAL CONSIDERATIONS

MOSHE TAL

Department of Life Sciences, Ben Gurion Universityof the Negev, P.O. Box 653, Beer Sheva 84105, Israel

(Received 25 February 1994; accepted 11 May 1994; editor T. A. Thorpe)

SUMMARY
In recent years attempts have been made to supplement traditionalbreeding for the production of salt-tolerantplants
with variabilityexistingin cellculture.The potentialcauses suggested as an explanation for the limitedsuccess of the in
vitroapproach include: a) lack, or loss during selection,of regeneration capability;b) the development of epigenetically
adapted cells;c) lack of correlationbetween the mechanisms of toleranceoperating in culturedcellsand mechanisms that
operate in cellsin the intactplant; and d) multigenicityof salttolerance.The recent successful production of healthy,
fertile,and geneticallystable salt-tolerantregenerants from cellsobtained from highly morphogenic explants which are
selected early in culture (using one-step or short-term strategies)for salttolerance,togetherwith the demonstration that
salt-sensitiveplantscan become tolerantby mutations in one or few genes, suggest that some of the potentiallimitations
can be overcome and that some of them may not exist at all.
Key words: salt tolerance; in vitro; selection; plants.

INTRODUCTION
The improvement of a plant's ability to produce economic yield
under conditions of abiotic stresses continues to be a challenge to
plant scientists, mainly in physiology and genetics. The genetical
research includes: a) traditional breeding of whole plants based on
morphologic (e.g., relative growth), physiologic (e.g., sodium accu-
mulation or exclusion), or molecular (e.g., RFLP and RAPD) selec-
tion criteria; b) in vitro selection in isolated tissues and cells; and c)
genetic engineering which includes the identification, characteriza-
tion, and manipulation of single gene sequences. In recent years
attempts have been made to supplement conventional breeding di-
rected toward the production of stress-resistant plants in general
and salt resistant in particular (Shannon, 1985; Tal, 1985; Epstein
and Rains, 1987; Blum, 1988; McNeilly, 1990; Forster, 1992,
1994; Galiba et al., 1992; Saranga et al., 1992) with variability
existing or produced in tissue and cell culture. The unique advan-
tages, the main methodologies, and accomplishments of the in vitro
production of salt-tolerant plants have been reviewed (Meredith,
1984; Chandler and Thorpe, 1986; Rains et al., 1986; Collin and
Dix, 1990; Nabors, 1990; Tal, 1990, 1993; Dix, 1993). Some of
these reviews also include short discussions of the potential causes
suggested as an explanation for the limited success of the in vitro
approach. These potential causes include: a) lack or loss during
selection of regeneration capability; b) the phenomenon of epigene-
tic adaptation; c) lack of correlation between the mechanisms of
tolerance operating in cultured cells and those of the whole plant;
and d) multigenicity of salt tolerance. However, recent successful
production of healthy, fertile, and genetically stable salt-tolerant
regenerants in various species (discussed by Dix, 1993 and Tal,
1993 and summarized in Table 1) suggested that some of the limita-
tions can be overcome and that some of them may not exist. Rea-
sons for this success may be: a) the use of explants with high mor-
phogenic potential which may ensure successful regeneration; b)
the use of one-step or short-term selection that may prevent the
development of epigenetically adapted cells which may obscure the
selection of rare mutants with true tolerance (Chandler and Vasil,
1984; McHughen and Swartz, 1984; Dix, 1993). The successful
production of salt-tolerant regenerants by one-step or short-term
selection suggests also that: a) at least some of the mechanisms of
salt tolerance operating in cultured cells operate also in cells of the
whole plant; and b) although the genetics of salt tolerance is com-
plex, the difference between salt-sensitive and -tolerant plant does
not necessarily require changes in many genes.
The present discussion concentrates mainly on existing data
which support the latter argument and, in addition, on the phenome-
non of epigenetic adaptation, and the correlation (or lack of) be-
tween mechanisms of tolerance which operate in cultured cells and
those operating in cells of the whole plant.
Multigenicity vs. monogenicity of salt tolerance. Whether the
difference between stress-sensitive and stress-tolerant genotypes is
necessarily based on many gene differences is one of the central
questions discussed in connection with the improvement of stress
resistance in general throughout genetic techniques. The genetics of
stress resistance in general and of salt tolerance in particular is
considered as complex (Norlyn and Epstein, 1984; Tal, 1984;
Blum, 1988). This complexity results because genes controlling
stress resistance in general are associated with mechanisms of adap-
tations that operate on different levels. McCue and Hanson (1990)

175
176 TAL

TABLE 1

IN VITRO SELECTION AND REGENERATION OF SALT-TOLERANTPLANTS: ONE-STEP OR SHORT-TERM STRATEGIES

Plant Explant Salt and conc. (g/liter) Selection Inheritance Reference

Rice Embryogenic
callus NaC1 (10, 20) One step Ro, R1, R2, R3 Vajrabhaya et al. (1989)
and R4
Sugarbeet Petioles (P) from in vitro NaC1(1, 2) Three cycles Ro, R1 Freytag et al. (1990)
multiplied shoots MgSO4 (2.0)
NaHCOs (1.6)
CaC12 (1.2)
CaSO4 (1.6)
Canola Highly morphogenic NaC1 (7.5-10) One step Ro, R1 Jain et al. (1990)
cotyledons
Canola Somaticembryos (SE) NaCI (up to 12.5) SE and Shoots Ro, R1 Kirti et al. (1991)
(twice) on salt
Flax Callus Sulfate salts (26.8) One step Ro, R1, R2 O'Connor et al. (1991)
(general vigor)
Alfalfa Callus newly initiated NaC1 (10.0) One step Ro, R1, R2 Winicov (1991)
from immature
ovaries
Coleus Leaves (disc) from in NaC1 (5.3) One step Ro, R1 Ibrahim et al. (1992)
vitro multiplied shoots (general vigor)
Tobacco Protoplasts (n) NaCI (11.7) Short term Ro • wild type Sumaryati et al. (1992)
X F1
BC1 F2
Vigna Cotyledonsfrom young NaCI (up to 150 Two steps Ro, R1 Gulati and Jaiwal (1993)
radiata in vitro-raised raM)
seedlings

classified the many types of adaptations evolved to drought and salt
stress in the whole plant as belonging to four levels: a) the develop-
mental level which includes the most complex mechanisms and
processes and therefore requires the interplay of products of many
genes. These processes are more easily related to agronomically
desirable traits because they are nearer to the output of the system
(Mabon, 1983). b) the structural level; c) the physiologic level; and
d) the biochemical or metabolic level which perhaps involve only
initial products of single genes, and are more easily related to sim-
ple genetic control and, therefore, lie within the reach of genetic
engineering (and, as we will see later, also of Mendelian genetics).
According to Cushman et al. (1990) the efficacy of the response by
four metabolic pathways (e.g., carbon metabolism, accumulation of
compatible osmolytes, sodium partition, and energy metabolism)
determines the tolerance phenotype.
Breeding for salt tolerance would be facilitated if the genes regu-
lating this characteristic could be identified. According to Cheese-
man (1988), based both on logic and on years of experience of
plant breeding, there will be no single gene or gene product which
determine "salt tolerance". He however agrees that absence of any
number of single genes may confer intolerance. General arguments
supporting the idea that single genes may have large effects on
stress resistance and that changes in the mean of the population
may occur via changes at a few or many loci have been discussed by
Tal (1984, 1985, 1990, 1993) and Hoffman and Parsons (1991).
Based on observations in cells and whole plants, Hasegawa et al.
(1990) and Amzallag et al. (1990) suggested that glycophytes and
halophytes both have the genetic make up that is required for the
existence of at least the cellular components of salt tolerance and
that the main difference between the two groups is in their ability,
which may depend on a relatively small number of regulatory genes,
to express this potential when required.
Although the expression of the various mechanisms of stress resis-
tance in general and of salt stress in particular depends on the
effects of several genes, a mutation in one of the key genes that
control these mechanisms may be enough to transfer a relatively
stress-resistant plant into a more sensitive one. Examples for such
effects include: 1) the mutation scabrous diminutive of pepper (Tal
and Benzioni, 1977) increases the sensitivity of the plant to NaCI by
decreasing its ability to exclude Na+ and to absorb K +. The mutant
is characterized also by changes in leaf anatomy (Tal et al., 1974)
and therefore may result from one mutation in a pleotropic gene or
(which is less likely) from two mutations in closely located genes. 2)
Single gene mutations that eliminate the first or both steps of the
biosynthetic pathway of glycinebetaine which was suggested as the
most effective compatible osmolyte in higher plants, were suggested
as a major cause for the relatively great sensitivity of rice cultivars
and some maize genotypes to drought and salt stress (Rathinasaba-
pathi et al., 1993). They suggested also that: a) the tribe Oryzeae
lost the ancestral ability to accumulate glyeinebetaine, which has a
central selective advantage under dry or saline conditions, during its
evolution in fresh water marshes; b) the genes for enzymes involved
in glycinebetaine biosynthesis would need to be introduced to im-
prove the drought- and salt-tolerance of, for example, tomato, po-
tato, and rice which do not accumulate glyeinebetaine but have
relatives that are accumulators of this osmolyte (MeCue and Han-
son, 1990). 3) Three mutants of the cyanobacterium Synechocystis
sp. PCC 6803 unable to tolerate high-salt concentrations were pro-
duced using random cartridge mutagenesis (Hagemann and Zuther,
1992). Salt sensitivity of one of the mutants was found to be caused
by a block in the biosynthesis of the osmolyte glucosylglycerol. No
physiologic defect responsible for the loss of salt tolerance could be
detected in the other two mutants. 4) Several wilty, drought-sensi-
tive, single-gene mutations have been isolated (probably because
 IN VITRO SELECTION FOR SALT TOLERANCE 177

TABLE 2

SINGLE-GENE MUTATIONS WHICH TRANSFORM THE PLANT FROM STRESS SENSITIVE TO STRESS RESISTANT
Plant Mutation(s) Resistance to Mechanism Reference

Soybean dominant NaC1 chloride exclusion Abel (1969)

Barley semidominant NaC1 high production of proline Kuehand Bright
(three alleles) (1982)
Arabidopsis recessive NaC1 high production of proline Lehleand Khan (1990)
Wheat ? frost ? Sutka and Shape
(1989)
Ceratopteris richardii two semidominant NaC1 ? Hickok et al. (1991)
(fern)
Alfalfa semidominant NaC1 ? Winicov (1991)
Wheat recessive osmotic stress turgor regulation by Morgan (1991)
osmotic adjustment
Yeast ? NaCI activity of Na+/H + Jia et al. (1992)
LiC1 antiperter
Nicotiana plumbaginifolia two dominant NaC1 and dehydration ? Sumaryati et al. (1992)
Grapevine dominant NaC1 chloride exclusion Sykes (1992)
Wheat two recessive" NaC1 sodium exclusion Taeb et al. (1992)
Arabidopsis three recessive NaC1 and osmotic stress ? Saleki et al. (1993)

~ Both genes, which are pleiotropic, are involved also in development (Vm and Ppa control vernalizationand photopefiod require-
ments, respectively).

they are easily recognized phenotypically) in higher plants. Various
causes for wiltiness have been described: abnormal xylem-vessel
development (Postlethwait and Nelson, 1957; Alldridge, 1964);
alteration in the ionic content of guard cells (Tal et al., 1976); low
osmotic pressure in the cell sap (Langridge, 1958); and deficiency
or insensitivity to abscisic acid [Tal and Nevo (1973)--in tomato;
Koornneef et al. (1982)--in Arabidopsis; Quarrie (1982)--in po-
tato; Wang et al. (1984)--in pea; Walker-Simmons et al. (1989)--
in barley; Parry et al. (1991)--in Nicotiana].
If transformation from relative resistance to sensitivity by one
gene mutation can happen, then why not the reverse, i.e., a change
from sensitive to resistant plant by one or a few mutations? Several
examples of single-gene mutational changes (four dominant, four
semi-dominant, and seven recessive) from stress sensitivity to resis-
tance have been reported in various species (Table 2). The mecha-
nisms responsible for the resistance are not known for most of the
mutations included in Table 2. However, such mutations as well as
those defective in some aspect of stress metabolism can be an excel-
lent tool to study the processes or mechanisms involved in the inte-
gration of response to salinity conditions (Cheeseman, 1988). Addi-
tional interested finding includes the inheritance of major compo-
nents of the mechanism of salt tolerance in citrus and grapevine. In
citrus Cl-ion and Na-ion exclusion seems to be inherited polygeni-
cally, whereas in grapevines Cl-ion exclusion may be expressed
either as a polygenic or monogenic trait depending on the parents
(Sykes, 1992). Forster et al. (1994) found a reduction in shoot
sodium content (which is considered as a central component of salt
tolerance in the Triticeae) in the barley cuhivar Golden Promise,
which is a gamma-induced mutation of the cultivar Maythorpe. They
suggested, therefore, that the ability to mutate genes involved in
salt-tolerance mechanisms clearly has profound implications for
adapting crop species to saline environments. Tolerances to several
abiotic stresses in the Triticeae were found to be influenced by
genes located on all seven homoeologous chromosome groups, from
which chromosomes 4 and 5 are predominants (Forster et al.,
1990). They suggested the possibility that genes involved in the
control of abiotic stresses are clustered, and that clustering of genes
controlling similar functions on specific chromosomes is a universal
phenomenon. Such closely linked genes that contribute to a single
character or to functionally related characters are designated as a
supergene (Futuyma, 1986). In accordance with some of the find-
ings related to the inheritance of salt tolerance, the results of all the
detailed genetic analyses of metal tolerance (which is simpler physio-
logically than salt tolerance in plants so far carried out have led to
the conclusion that there are dominant major genes for tolerance
with other genes with small effects (modifiers) present as well (Mac-
hair et al., 1992).
Molecular biology provides tools for dissecting the tolerant pheno-
type in a reductionist manner (Cushman et al., 1990). The progress
made in the molecular study of gene expression in relation to salt
tolerance as well as problems and perspectives have been discussed
in recent reviews (Cushman, 1990; Scandalios, 1990; Hurkman,
1992; Winicov, 1993). Winicov (1993) discussed recent promising
achievements in the molecular manipulation of genes involved in the
metabolism of osmolytes. It includes the first molecular demonstra-
tion of successful transformation from salt sensitivity to tolerance by
the engineering of a single gene. A transgenic tobacco plant to
which a bacterial gene mtl D (which encodes the enzyme mannitol-
1-phosphate dehydrogenase) was engineered, acquired the ability
to accumulate mannitol in roots and leaves and to tolerate high
salinity (Tarczynski et al., 1993).
Epigenetic adaptation. "Adaptation" will be used here to de-
fine stable epigenetic alterations that are inherited only through
mitosis and not through meiosis. According to McHughen and
Swartz (1984) and Chandler and Vasil (1984), gradual exposure of
cells to salt stress is ineffective for the selection of salt-tolerant
mutants, because non-tolerant cells can quite readily undergo an
epigenetic adaptation to increasing levels of salinity and thus ob-
scure the selection of rare mutants with true, i.e., meiotically in-
herited, tolerance (Dix, 1993). It has been recommended that adap-
tation can be prevented by exposing the cells directly to sublethal
salt concentrations (McHughen and Swartz, 1984; Blum, 1988;
178
Sumaryati et al., 1992). The knowledge of adaptation and the mech-
anisms controlling its occurrence may enable a distinction between
mutant and adapted cells and thus may help to improve the effi-
ciency of selection of salt-tolerant mutant cells (Tal, 1993). Based
on a number of studies of methylation pattern in cultured cells, Ball
(1990) suggested that altered methylation can provide an explana-
tion for epigenetic changes that are sometimes observed in tissue
euhure experiments.
The possibility that adaptation, which is an epigenetic phenome-
non, depends on changes in the pattern of DNA methylation (which
is also, at least in part, epigenetic) that affects gene expression is
being studied in our laboratory (Sabbah et al., in preparation). A
higher percentage of 5-methylcytosine was found in the DNA (in-
eluding gross DNA and DNA isolated from DNaseI-sensitive chro-
matin) of NaCl-adapted cultured cells of potato as compared with
that of control cells. This difference in methylation between the two
cell types was much more noticeable in the DNA isolated from
DNaseI-sensitive chromatin which presumably represents regions
with (relatively to insensitive chromation) more active genes
(Spiker, 1985). Sabbah et al. (in preparation) found also that the
level of some electrophoretic protein bands increased and some
decreased in the adapted cells as compared with the control ones.
To verify the possibility that adaptation to NaCI and methylation of
DNA are causally related, it is planned to study DNA methylation in
sequences of specific salt responsive genes, using comparative di-
gestion of DNA with methylation-sensitive (isoschizomers) restric-
tion enzymes.
Are mechanisms of salt tolerance operating in cultured cells corm-
lated with those operating in cells in the whole plant? Lack of
positive correlation between the mechanisms of tolerance operating
in isolated cultured cells and those operating in the intact plant has
been suggested as one of the main reasons for the limited success of
the in vitro selection of salt-tolerant plants (Lebrun et al., 1985;
Vasil, personal communication; Dracup, 1991). According to Dra-
cup (1991) increased levels of salt tolerance in cultured cells tend
not to be expressed in regenerated plants for the following reasons:
a) many responses to high NaC1 are associated with the integrated
functioning of the whole plant, rather than merely cellular responses
to high NaC1; b) the hormonal and nutritional (an abundant supply
of sugars which can be used as cheap osmotica rather than the salt
ions, for example) environments differ between cells in culture and
in the whole plant: traits selected in cultured cells, therefore, may
not be expressed uniformly throughout the plant because expres-
sion may depend on variable hormonal and nutritional environ-
ments. It should be added that cells in culture and the regenerant
plants are usually exposed to completely different physical (tempera-
ture, light, and humidity) external conditions.
The recent successful production of salt-tolerant regenerants
means that, although there is a large difference between the condi-
tions where the cultured cells are selected and the conditions where
the salt tolerance of the regenerants is evaluated, at least some of
the cellular mechanisms of salt tolerance operating in cultured cells
operate also in the regeneranted plants.
A commonly used test for the existence of a correlation between
the mechanisms operating in cultured cells and in the whole plant is
based usually on the comparison of salt tolerance in the whole plant
and in cells isolated from it. All possible correlations were found
between the response to salt of the whole plant and the response of
cells in culture (summarized by Tal, 1990). A positive correlation
TAL
where the whole plant and the cultured cells are both tolerant o:
sensitive to salt is interpreted as an indication for the operation o:
lack of operation of similar cellular mechanisms on both levels. 1
negative correlation where the whole plant is salt tolerant and th,
isolated cells are sensitive was regarded as an indication for th,
operation of mechanisms depending on the organization of the cell..
in tissues in the whole plant. An opposite negative correlation
where the plant is sensitive and the isolated cells are resistant, wa.,
suggested (Flowers et al., 1985) to result from the different condi
tions surrounding the cells in the leaf and in culture: although effec
rive salt exclusion at the cellular level is advantageous in vitr,
(where carbohydrate supply should be unlimited) it can be disad
vantageous for leaf cells in vivo, for which carbohydrate supply
not unlimited and ions excluded from leaf cells and accumulated it
the apoplast may induce water deficit and loss of turgor (Oertli
1968). Although the salt tolerance of the cultured cells and of th,
intact plants (from which the cells are derived) is usually evaluatec
under very different conditions, such comparative studies sug
gested, according to Dix (1993), that there was frequently, but no
always, a sufficient cellular basis for the differences in salinity-toler
ant mutants to justify in vitro selection as an approach to obtair
tolerant mutants.
CONCLUSION
The recent successful production of healthy, fertile, and geneti
tally stable salt-tolerant regenerants in various species (Table 1
can be the basis for a renewed optimism in the application of the ir
vitro approach for the selection of salt-tolerant plants. The demon.
stration of several eases of single-gene mutations which can trans.
form salt-or stress-sensitive plants to tolerant ones (Table 2) add.,
strength to this argument.
Until recently, information about the inheritance of salt toleranc~
in regenerated plants was limited to two tobacco examples wberc
the tolerant cells were obtained by selection performed over ar
extended period (Nabors, 1983; Bressan et al., 1985). In these two
cases the inheritance was found to be complex and not strictly Men-
delian. However, two cases of single-gene mutational changes frorr
salt sensitivity to tolerance were reported recently in regenerant,.
produced from cells obtained by short-term selection. It is expectec
that similar simple genetic changes characterize the production o~
the other salt-tolerant regenerants from cells obtained by the short.
term selection (Table 1). Such mutations as well as those defectiv~
in some aspect of stress metabolism can be an excellent tool tc
study the genetic and physiologic processes and mechanisms that
are responsible for the integration of response to stress conditions
Some of these mutations (Table 2) are involved in the exclusion oJ
CI or Na from the shoot, which was suggested as an effective selec.
tion criterion for salt tolerance in various species (discussed b)
Noble and Rogers, 1992).
REFERENCES
Abel, G. H. Inheritance of the capacityfor chloride inclusion and exclusior
by soybean. Crop. Sei. 9:697-698; 1969.
Alldridge, N. A. Anomalousvessel elements in wihydwarftomato. Bot. Gaz.
125:138-142; 1964.
Amzallag, G. N.; Lerner, H. R.; Poljakoff-Mayber, A. Induction of in-
creased salt tolerance in Sorghum bicolor by NaCIpretreatment. J.
Exp. Bot. 71:29-34; 1990.
Ball, S. G. Molecular basis of somaelonal variation. In: Bajaj, Y. P. S., ed.
 IN VITRO SELECTION FOR SALT TOLERANCE
Biotechnology in agriculture and forestry, vol. 11. Somaclonal varia-
tion in crop improvements. Berlin: Springer-Verlag; 1990:
134-152.
Blum, A. Plant breeding for stress environments. Boca Raton, FL: CRC
Press; 1988.
Bressan, R. A.; Singh, N. K.; Handa, A. K., et al. Stable and unstable
tolerance to NaCI in cultured tobacco cells. In: Freeling, M., ed.
Plant genetics: proceedings of the third annum ARCO plant cell
research institute--UCLA symposium on plant biology. New York:
A. R. Liss; 1985:755-769.
Chandler, S. F.; Thorpe, T. A. Variation from plant tissue cultures: biotech-
nological application to improving salinity tolerance. Biotechnol.
Adv. 4:117-135; 1986.
Chandler, S. F.; Vasil, I. K. Selection and characterization of NaC1 tolerant
cells from embryogenic cultures of Pennisetam purpureum Schum.
(Napir grass). Plant Sci. Leu. 37:157-164; 1984.
Cheeseman, J. M. Mechanisms of salinity tolerance in plants. Plant Physiol.
87:547-550; 1988.
Collin, H. A.; Dix, P. J. Culture systems and selection procedures. In: Dix,
P. J., ed. Plant cell line selection procedures and applications. New
York: VCH Weinheim; 1990:3-18.
Cushman, J. C.; DeRocher, E. J.; Bohnert, H. J. Gene expression during
adaptation to salt stress. In: Katterman, F. R., ed. Environmental
injury to plants. San Diego, CA: Academic Press; 1990:
173-203.
Dix, P. J. The role of mutant cell lines in studies on environmental stress
tolerance: an assessment. Plant J. 3:309-313; 1993.
Dracup, M. Increasing salt tolerance of plants through cell culture requires
greater understanding of tolerance mechanisms. Anst. J. Plant Phys-
iol. 18:1-15; 1991.
Epstein, E.; Rains, D. W. Advances in salt tolerance. Plant Soil 99:17-29;
1987.
Flowers, T. J.; Lachno, D. R.; Flowers, S. A., et al. Some effects of sodium
chloride on cells of rice cultured in vitro. Plant Sci. Lett. 39:205-
211; 1985.
Forster, B. P. Genetic engineering for stress tolerance in the Triticeae.
Prec. R. Soc. Edinb. 99B:89-106; 1992.
Forster, B. P.; Phillips, M. S.; Miller, T. E., et al. Chromosome location of
genes controlling tolerance to salt (NaCI) and vigour in Hordeum
vulgare and H. chilense. Heredity 65:99-107; 1990.
Forster, B. P.; Pakniyat, H.; Macaulay, W., et al. Variation in the leaf
sodium content of the Hordeum vulgare (barley) cultivar Maythorpe
and its derived mutant cv. Golden Promise. Heredity 73:249-253;
1994.
Freytag, A. M.; Wrather, J. A.; Erichsen, A. W. Salt tolerance sugarbeet
progeny from tissue cultures chalhnged with multiple salts. Plant
Cell Rap. 8:647-650; 1990.
Futuyma, D. J. Evolutionary biology, 2od ed. Sunderland, MA: Sinauer
Associates, Inc. Publishers; 1986.
Galiba, G.; Simon-Sarkadi, L.; Kocsy, G., et al. Possible chromosomal
location of genes determining the osmoregulation of wheat. Theor.
Appl. Genet. 85:415-418; 1992.
Gulati, A.; Jaiwal, P. K. Selection and characterization of mannitol-tolerant
callus lines of V/gna rad/ata (L.) Wilezak. Plant Cell Tissue Organ
Cult. 34:35-41; 1993.
Hagemann, M.; Zuther, E. Selection and characterization of mutants of
Cyanobacterium Syneehocystis sp. PCC 6803 unable to tolerate
high salt concentrations. Arch. Mierobiol. 158:429-434; 1992.
Hasegewa, P. M.; Binzel, M. L.; Reuveni, M., et al. Physiological and
molecular mechanisms of ion accumulations and compartmenta-
tions contributing to salt adaptation of plant cells. In: Bennett, A. B.;
O'Neill, S. D., eds. Horticultural bioteehnology. New York: Wiley-
Liss; 1990:295-304.
Hickok, L. G.; Vogelien, D. L.; Varne, T. R. Selection of a mutation confer-
ring high NaCI tolerance to gametophytes of Ceratopteris. Theor.
Appl. Genet. 81:293-300; 1991.
Hoffmann, A. A.; Parsons, P. A. Evolutionary genetics and environmental
stress. Oxford, England: Oxford Science Publications; 1991.
Hurkman, W. J. Effect of salt stress on plant gene expression: a review.
Plant Soil 46:145-151; 1992.
Ibrahim, K. M.; Collins, J. C.; Coilin, H. A. Characterization of progeny of
179
Coleus blumei following an in vitro selection for salt tolerance. Plant
Cell Tissue Organ Cult. 28:139-145; 1992.
Jia, Z. P.; McCullough, N.; Martel, R., et al. Gene amplification at a locus
encoding a putative Na+/H + antiporter confers sodium and lithium
tolerance in fission yeast. EMBO J. 11:1631-1640; 1992.
Jain, R. K.; Jain, S.; Nainawatee, H. S., et al. Salt-tolerance in Brassica
juncea L. I. In vitro selection, agronomic evaluation and genetic
stability. Euphytica 48:141-152; 1990.
Kirti, P. B.; Hadi, S.; Kumar, P. A., et al. Production of sodium-chloride-tul-
erant Brassica juncea plants by in vitro selection at the somatic
embryo level. Theor. Appl. Genet. 83:233-237; 1991.
Koornneef, M.; Jorna, M. L.; Brinkhorst-van der Swan, D. L. C., et al. The
isolation of abseisic acid (ABA) deficient mutants by selection of
induced revertants in non-germinating gibberelin sensitive lines of
Arabidopsis thaliana (L.) Heyah. Theor. Appl. Genet. 61:
385-393; 1982.
Kueh, J. S. H.; Bright, S. W. J. Biochemical and genetical analyses of three
proline-accumulating barley mutants. Plant Sci. Lett. 27:
233-241; 1982.
Langfidge, J. An osmotic mutant ofArabidopsis thaliana. Aust. J. Biol. Sci.
11:457-470; 1958.
Lebrun, L.; Rajasekaran, K.; Mullins, M. G. Selection in vitro for NaCl-tol-
erance in Vitis rupestris Scheele. Ann. Bot. 56:733-739; 1985.
Lehle, F. R.; Kahn, R. A. Arabidopsis mutant with enhanced NaC1 toler-
ance. In: Schweizer, D.; Peuker, K.; Loidl, J., eds. Fourth Intern.
Conf. Arabidopsis Research, Inst. Cytology and Genetics, University
of Vienna, Vienna, 1990. [Published by the meeting organizers.]
Lu, D. B.; Scars, R. G.; Paulsen, G. M. Increasing stress resistance by in
vitro selection for abscisic acid insensitivity in wheat. Crop. Sci.
29:939-943; 1989.
Macnair, M. R.; Cumbs, Q. J.; Meharg, A. A. The genetics of arsenate
tolerance in Yorkshire fog, Holcus /anatus L. Heredity 69:325-
335; 1992.
Mabon, J. D. Limitations to the use of physiological variability in plant
breeding. Can. J. Plant Sci. 63:11-21; 1983.
McCue, K. F.; Hanson, A. D. Drought and salt tolerance: towards under-
standing and application. Trends Biotech. 8:358-362; 1990.
McHugen, A.; Swartz, M. A tissue culture derived salt-tolerant line of flax
(Linum usitatissimum). J. Plant Physiol. 117:109-117; 1984.
McNeilly, T. Selection and breeding for salinity tolerance in cross species.
A case for optimism? Acta Oecol. 11:595-610; 1990.
Meredith, C. P. Selecting better crops from cultured cells. In: Gustafson,
J. P., ed. Gene manipulation in plant improvement. 16th Stadler
Genetics Symposium. New York: Plenum Press; 1984:503-528.
Morgan, J. M. A gene controlling differences in osmoregulation in wheat.
Aust. J. Plant Physiol. 18:249-257; 1991.
Nabors, M. W. Increasing salt and drought tolerance of crop plants. In:
Randall, D. D.; Blevins, D. B.; Larson, R. L., et al., eds. Current
topics in plant biochemistry and physiology, vol. 2. Columbia, MO:
University of Missouri; 1983:167-186.
Nabors, M. W. Environmental stress resistance. In: Dix, P. J., ed. Plant cell
line selection procedures and applications. New York: VCH Wein-
heim; 1990:167-186.
Noble, C. L.; Rogers, M. E. Arguments for the use of physiological criteria
for improving the salt tolerance in crops. Plant Soil 146:99-107;
1992.
Norlyn, J. D.; Epstein, E. Variability in salt tolerance of four Triticah lines
at germination and emergence. Crop. Sci. 24:1090-1092; 1984.
O'Connor, B. J.; Robertson, A. J.; Gusta, L. V. Differential stress tolerance
and cross adaptation in a somaclonal variant of flax. J. Plant Physiol.
139:32-36; 1991.
Oertli, J. J. Extracellular salt accumulation, a possible mechanism of salt
injury in plants. Agrochemica 12:461-469; 1968.
Parry, A. D.; Blonstein, A. D.; Babiano, M. J., et al. Abscisic-acid metabo-
lism in a wihy mutant of Nicotiana plumbaginifolia. Planta
183:237-243; 1991.
Pesththwait, S. N.; Nelson, O. E. A chemically wilted mutant of maize. Am.
J. Bot. 44:628-633; 1957.
Quarrie, S. A. Droopy: a wilty mutant of potato deficient in abseisic acid.
Plant Cell Environ. 5:23-26; 1982.
Rains, D. W.; Croughan, S. S.; Croughan, T. P. Isolation and characteriza-
180
tion of mutant cell lines and plants: salt tolerance. In: Vasil, I. K.,
ed. Cell culture and somatic cell genetics of plants, vol. 3. Orlando,
FL: Academic Press; 1986:537-547.
Rathinasnhapathi, B.; Gage, D. A.; Mackill, D. J., et al. Cultivated and wild
species do not accumulate glycinehitalne due to defliciencies in two
biosynthetic steps. Crop. Sei. 33:534-538; 1993.
Saleki, R.; Young, P. G.; Lefebvre, D. D. Mutants of Arabidopsis thaliana
capable of germination under saline conditions. Plant Physiol.
101:839-845; 1993.
Saranga, Y.; Cahaner, A.; Zamir, D., et al. Breeding tomatoes for salt
tolerance: inheritance of salt tolerance and related traits in interspe-
cific population. Theor Appl. Genet. 84:390-396; 1992.
Scandalies, I. G., ed. Genomic responses to environmental stress: advances
in genetics, vol. 28. San Diego, CA: Academic Press; 1990.
Shannon, M. C. Principles and strategies in breeding for high salt tolerance.
Plant Soil 89:227-241; 1985.
Spiker, S. Plant chromatin structure. Ann. Rev. Plant Physiol. 36:235-
253; 1985.
Sumaryati, S.; Negrntin, I.; Jacobs, M. Characterization and regeneration of
salt- and water-stress mutants from protoplast culture of Nicotiana
plumlmginifolia (Viviani).Theor. Appl. Genet. 83:613-619; 1992.
Sutka, J.; Snape, J. W. Location of a gene for frost resistance on chromo-
some 5A of wheat. Euphytica 42:41-44; 1989.
Sykes, S. R. The inheritance of salt exclusion in woody perennial fruit
species. Plant Soil 146:123-129; 1992.
Tacb, M.; Koebner, R. M. D.; Forster, B. P., et al. Association between
genes controlling flowering time and shoot sodium accumulation in
the Triticeae. Plant Soil 146:117-121; 1992.
Tal, M. Physiological genetics of salt resistance in higher plants: studies on
the level of the whole plant and isolated organs, tissues and cells. In:
Staples, R. C.; Toenniessen, G. H., eds. Salinity tolerance on plants.
Strategies for crop improvement. New York: Wiley & Sons;
1984:301-320.
Tal, M. Genetics of salt tolerance in higher plants: theoretical and practical
considerations. Plant Soil 89:199-226; 1985.
TAL
Tal, M. Somaclonal variation for salt resistance. In: Bajaj, Y. P. S., ed.
Biotechnology in agriculture and forestry, vol. 11. Somaclonal varia-
tion in crop improvement. Berlin: Springer-Verlag; 1990:
236-257.
Tal, M. In vitro methodology for increasing salt tolerance in crop plants.
Acta Hortic. 336:69-79; 1993.
Tal, M.; Benzioni, A. Ion imbalance in Capsicum annuum, Scabrous dimin-
utive, a wihy mutant of pepper. I. Sodium fluxes. J. Exp. Bot.
28:1337-2341; 1977.
Tal, M.; Nevo, Y. Abnormal stomatal behavior and root resistance, and
hormonal imbalance in three wihy mutants of tomato. Biochem.
Genet. 8:291-300; 1973.
Tal, M.; Witztum, A.; Shifriess, C. Abnormal stomatal behavior and leaf
anatomy in Capsicum annuum, scabrous diminutive, a wilty mutant
of pepper. Ann.Bot. 38:983-988; 1974.
Tal, M.; Esbel, A.; Witztum, A. Abnormal stomatal behaviour and ion imbal-
ance in Capsicum scabrous diminutive. J. Exp. Bot. 27:953-960;
1976.
Tarczynski, M. C.; Jensen, R. G.; Bohnert, H. J. Stress protection of trans-
genic tobacco by production of the osmolyte, mannitol. Science
259:508-510; 1993.
Vajrabhaya, M.; Thanapaisai, T.; Vajrabhaya, T. Development of salt taler-
ant lines of KDML and LPT rice cultivars through tissue culture.
Plant Cell Rep. 8:411-414; 1989.
Walker-Simmons, M.; Kudrna, D. A.; Warner, R. L. Reduced accumula-
tion of ABA during water stress in a molybdenum cofactor mutant of
barley. Plant Physiol. 90:728-733; 1989.
Wang, T. L.; Donkin, M. E.; Martin, E. S. The physiology of a wihy pea:
abscisic acid production under water stress. J. Exp. Bot 35:1222-
1232; 1984.
Winicov, I. Characterization of salt tolerant alfalfa (Medicago sativa L.)
plants regenerated from salt tolerant cell lines. Plant Ceil Rep.
10:561-564; 1991.
Winicov, I. Gene expression in relation to salt tolerance. In: Basra, A. S.,
ed. Stress-induced gene expression in plants. Harwood: Academic
Publishers; 1993:61-85.