BREEDING OF ABIOTIC STRESSES IN VEGETABLE CROPS

Presented by K. Indira VHM/10-09

Every plant required some physical and chemical factors in optimum amount at a right time for its proper growth and development. Any deviation from this optimal condition of any factor essential for its growth will lead to aberrant change in physiological processes and due to this plant body will experience tension and this state can be referred as plant under stress. Stress caused by environmental factors such as drought or extreme heat or cold or by deficiency or toxicity of minerals is called as abiotic stress.

Environmental factors responsible for abiotic stress in vegetable crops:

Drought Excessive moisture High and low temperatures Salinity Alkalinity Atmospheric pollutes

Among all those factors responsible for abiotic strees the most important factors limiting productivity are drought, heat stress and mineral stress. About 20% of the land is limited by mineral stress, 26% by drought stress and 15% by freezing stress (N. K. Srinivasa Rao and R. M. Bhatt). It has been estimated that only 10% of the worlds arable land may be classified in a non stress category.

Breeding methods
a. Conventional methods:
1. Selection 2. Recurrent selection 3. Pedigree method 4. Back cross method 5. Bulk pedigree method 6. Mass selection 7. Mutation breeding 8. Interspecific hybridization

b. Non conventional methods:

1. Tissue culture techniques 2. Molecular/ genetic marker based approaches 3. Genetic engineering techniques

Choice of breeding method mainly depend on Mode of reproduction Mode of gene action Source of resistance varieties available Priority assigned to goal

Screening
Lewis and Christiansen advocated following three methods of testing breeding material for stress resistance. 1. Indirect selection:- Materials are screened under natural stress environment conditions and a variety performing well under such stress conditions may be considered stress tolerant. 2.Direct selection:- In this case , materials are grown under uniform stress condition for screening. This is growing of tomato in winter and summer for screening of fruit set at low and high temperatures, respectively. 3.Controlled condition:- The most precise accurate method is to test the breeding material under controlled conditions.

A.Drought tolerance: Drought is the important environmental constraint that limits the expression of genetic potential of many crops. Drought is the inadequacy of water availability, including precipitation and soil moisture storage capacity, in quantity and distribution during the life cycle of a crop to restrict the expression of its full genetic yield potential. Plants adapt to drought by avoidance or tolerance mechanisms. The ability of the plant to with stand low tissue water potentials may be considered as drought tolerance. The ability of the plant to maintain favorable internal water balance under stress is considered as drought avoidance.

Classification of drought stress

Type of stress Mild stress Moderate stress severe stress W (Mpa) 0.1 MPa - 1.2 to -1.5 MPa < -1.5MPa Reduction in RWC 8 -10% >10 to < 20% >20%

Hsiao (1973) Drought tolerance is achieved by the involvement and coordination of several physiological attributes during the limited water supply.

Osmotic adjustment: It is the process in which organic solutes accumulate and maintain the turgor of cells during limited water supply. It can play a major role in determining the drought resistance by a) maintaining turgor over fluctuating soil water potentials, b) maintaining stomatal conductance and thus photosynthesis. Various compatible solutes like sugars, organic acids, amino acids, sugar alcohols or ions are responsible for osmotic adjustment.

Great genotypic variability exists in vegetable crops for osmotic adjustment. Genotypes with high osmotic adjustment have high dehydration tolerance and senescence may also be delayed. Srinivasa rao and Bhatt reported, tomato cultivars Arka Saurabh, Pusa Early Dwarf and Sioux have better osmotic adjustment and thus relatively higher yield in these cultivars under stress conditions.

Hormonal accumulation: Water deficits exert a profound effect on hormonal distribution, cytokinins and abscisic acid (ABA). As a consequence of water deficit, cytokinin activity is reduced and ABA is increased. An increase in the levels of ABA reduces stomatal opening, while cytokinins exerts an opposite effect. The ABA content was higher in cv. Arka Saurabh as compared to cv. Sel.1 of tomato. The content of ABA was almost three times higher in Arka Saurabh, a moderately stress tolerant cultivar, as compared to Sel-1, a sensitive cultivar under water stress.

Proline accumulation: Various metabolites are accumulated in the plant tissues upon reduction in leaf water status. Of these, free proline amino acid accumulated at very high concentrations with a rapid decrease in concentration upon rehydration (Srinivasa Rao, 1986). Proline accumulation have been shown to be adaptive mechanism of stress tolerance. But conflicting results claim a positive or a negative association with drought resistance. Significant accumulation of stress induced proline was studied in 8 cultivars of tomato at 0, 4, 7 and 10 days of stress (Thakur, 1991). High proline accumulating cultivars, Rupali, Goldmaker, Vaishali and KT- 10 were found to be more susceptible to imposed levels of water stress.

Drought resistance is measured by attributes like germination % of seed, rate of seedling growth, stomatal control of transpiration, proline content and yield under drought conditions. Great variation is there in the ability of genotypes to tolerant different degrees of drought. Generally, the direct selection method is used to identify resistant accessions. In case of indirect selection method, tolerant accessions are identified on the basis of high yield under drought conditions.

Drought resistance in tomato was assessed on the basis of % of seed germination in sucrose solution at different concentrations. The germination % of highly resistant varieties was 65% in sucrose solution, where in other varieties, germination % was less than 65% (Boyarshinova, G.A., 1981). In tomato, Strain B and Pritchard exhibited the highest germination and dry weight at the highest moisture stress. L. pennellii has been reported to be the prime example of drought resistance on the basis of plant growth.

Saladette and two accession of L..esculentum var. cerasiforme were found tolerant to moisture stress up to 10 days on the basis of growth rate and leaf area parameters. The red rock tomato variety grows well under dry conditions, which is probably due to a deep tap root system (Stoner,A.K., 1978). In soybean, leaf-specific weight was the most important parameter for drought stress resistance. In Vigna unguiculata, selections from, CB 5 exhibited greater drought stress resistance, earlier flowering and exhibited pod development at early stage than other varieties (Hall,A.E., and Grantz, D.A., 1981).

IGFRI 450 and IGFRI 437 were found to be most tolerant varieties. In Phaseolus vulgaris, Pinto and white half runner, and an accession of P. acutifolius were grown under drought stress conditions. Proline content is the indirect method of assessing drought resistance. Low proline accumulation was associated with drought resistance. In P. vulgaris, varieties likeMoruna and Aroana performed better under drought stress and accumulated less proline.

Experiments conducted in water melon under desert zone conditions concluded that drought resistance is associated with a reduced transpiration surface, high water retention, and closed stomata at high temperature (Malinina,M.I., 1971). Varieties of an extremely early type may escape drought stress.

Excessive moisture tolerance: The nature of excessive moisture damage depends upon crop species, varieties, age of the plant and type of soil. It reduces oxygen supply to the plant roots which results in an accumulation of endogenous ethylene in the plant. Under water logged condition the hormonal balance between root and shoot becomes upset. In tomato, flooding drastically reduced the GA level in the xylem sap of the plant (Reid,D.M., Crozier,A., Harvey, B.M.R., 1969).

Genotypic variation for susceptibility to flooding injury has been noticed in lycopersicon. A total of 4630 accessions were screened and flood tolerance was maximum in 8 accessions i.e ,lycopersicon esculentum L-123, L-125, L-973, L-3072, L-3091, L4313, L4360 and L. pimpinellifolium L-4422. In Tomato, Nagcarlan a primitive cultivar from Philippine LA 1421, member of the cerasiforme has been found tolerant to excessive moisture.

In soybean, 20 varieties were screened under flooded conditions, of which six varieties were comparatively superior for yield under water logged conditions (Matsukawa,I., Tanimura, Y.., and Banba,H..,). Varieties resistant to excessive moisture also showed the most rapid conversion of ethylene to ethylene oxide. In Vicia faba, water logged susceptible varieties yield measurable amount of ethylene oxide(Dodds, J.H.,Smith,A.R., and Hall,M.A.,).

Temperature tolerance
Heat tolerance: Heat tolerance is measured by the ability of the plant to grow under high temperature conditions. Heat resistance is a genetically controlled attribute. Heat tolerance has been studied in Solanum tuberosum (Khurana, S.C., and Terekhina, L.N., 1982). Potato leaf cuttings are used for heat tolerance studies in screening technique. Wild and cultivated species have been tested for heat and frost resistance. S. tuberosum was more heat tolerant than wild species.

Recurrent selection has been used in potato for developing a heat tolerant variety (Gautney, T.L., and Haynes, F.L., 1983). In tomato fruit set due to high temperatures is a general problem in tropical and sub tropical countries. When the temperature exceeds 32C, the fruit set is reduced. In Lycopersicon esculentum, heat tolerance is measured by the ability of a plant to set the fruits at a high temperature. Adverse effect of high temperature on fruit set is of extensively studied at 40C, ovule abortion and endosperm degeneration was observed by iwahori

There is a great variation among the tomato accessions in the ability to set fruits at high temperatures. Variety Hs-102 had the ability to set fruits in April when temperature is about 35 to 38C. However no accession set fruit in may- June. BL-6807, a cold set selection, had the highest ability to set fruit when pollen was exposed to high temperature. Saladette developed by Paul leeper in Texas, is heat tolerant. UC-82B is comparatively less heat tolerant than Saladette. The heat tolerance increased much faster in Saladette than in UC-82B after exposure to 35C.

In Saladette, there was normal endothecium formation in anthers at high temperatures which is essential for stamen and pollen thecae opening, which helps in pollination. Six varieties have been studied for pollen weight per flower which is drastically reduced at high temperatures. The maximum quantity of pollen was found in variety saladette. Heat tolerance exhibited low heritability and was highly influenced by environmental factors, thus indicating less scope for selection. Mass and pedigree methods are used in tomato.

In Phaseolus vulgaris, high temperatures drastically reduced the number of pods. Among Phaseolus species, Phaseolus coccineus was more sensitive and P. vulgaris was heat tolerant . The tolerance is controlled by few genes. Narrow sense heritability ranged from 2.9 to24%.

In Vigna unguiculata, abscission of flowers at high temperatures is very common. Indehiscence of the anther and low pollen viability are the main reasons for flower abscission. Screening studies have indicated that the genotypes- Tvu 4552, PI 204647, and prima set pods at high temperatures whereas, the other 55 genotypes flowered with none or a few pods produced.

Cold tolerance: A temperature of 0C or below causes severe damage in tropical and sub tropical crops like tomato, egg plant, cucurbits, okra and beans. Low temperature cause injury mainly to germination, growth, fruit set and yield when seed or plant of a chill sensitive crop is exposed to chilling temperatures. Potato, tomato, bean and cucurbits do not resist temperatures below -2 to -3C and exhibits frost injury symptoms. Whereas cole crops, spinach and peas can tolerate frost up to -3 to -5C. Cold tolerance is measured by the ability to germinate the seed, to set fruit at low temperatures and to withstand chilling temperatures

Low temperature germination: Optimum temperature required for uniform quick germination is about 15 to 25C for most of the vegetable crops. If temperature goes down to 15C there is a reduction in seed germination and it is drastically reduced at 10C or below. Low temperature seed germination is a heritable characteristic and was controlled by additive gene action. In Lycopersicon, it is mostly controlled by recessive factors and in muskmelon by dominant factors

The seeds of tomato genotypes widely differ in their ability to germinate at a low temperature. Eight accessions, including five wild types, were screened at low temperatures. One accession of L. Chilense LA 460 and one accession of L. Peruvianum PI 126435 germinated rapidly at 10C (Scott, S.J., and Jones, R.A., 1983). In Hisar, the temperatures in winter is very low and there is a problem of seed germination. All the germplasm were screened and only L.hirstum f.glabratum germinated and grew well under such conditions

There is a significant variation among the varieties of cucumber for germination from 15 to 35 C. 19 varieties, 8 breeding lines, and 176 introductions were screened, of which 5 varieties germinated at moderately low temperature (Wehner, T.C., 1982). Over 7000 accessions of cucumis melo were screened for germination and development at 15C, of these, 4 were found promising ( Hutton,M.G. and Loy, J.B., 1982)

Mass selection and recurrent selection can be followed for the improvement of germination at low temperatures. The ability to germinate at low temperatures was improved when mass selection was followed for 4 years in cucumber (Schulte, H.K., and Grote, U., 1974). Nienhuis et al. concluded that recurrent selection was effective in improving the germination % at suboptimal temperatures (15C) in cucumber.

Low temperature fruit set: Cold tolerance is also measured by the ability to set fruits under low temperature conditions. The normal temperatures for fruit set is from 15 to 25C, depending upon the variety. A temperature below 10 C limits the fruit set. Poor fruit set at low temperature is due to poor anther dehiscence, poor pollination, poor pollen viability and slow pollen tube growth. Hence artificial pollination is required for good fruit set.

In Hisar, maximum fruit set has been recorded at January in an accession of lycopersicon pimpinellifolium and now it is used to develop a cold tolerant variety (Kalloo and Banerjee, M.K., 1970). Under Hisar conditions, varieties like Cold set, Ostenkinskiz, Montfavet, and Precoce, exhibited better fruit set in winter. Cold set and Precoce varieties have good fruit set but because of their small fruit size they have less economic value. Hence these can be used only in hybridization. A parthenocarpic variety may have better scope for fruit set under low temperature conditions which is controlled by pat gene. Severianin exhibits facultative parthenocarpy and is used in cold-resistant breeding program.

Chilling tolerance: L. esculentum is highly sensitive to chilling temperatures . If temperature goes down to10C the growth of the plant almost ceases. For optimum growth, an 18C night temperature is required. L. hirsutum from high elevation can tolerate as low temperature as 6C. The seedling of L. hirstum, LA 1363, developed the first true leaf even at 0 C and a day temp of 20C, where as L.esculentum variety failed to develop its first true leaf under these conditions. The chilling resistance of L.hirsutum is heritable and can be transferred to L.esculentum (B.D. Patterson, R. Paull and R.M. Smillie , 1978).

In carrot, chantenay, group roots, less ice was formed and phloem and the skin was rarely spilt by ice formation. In celery, winter hardiness is studied in red and white commercial varieties. Red varieties showed better survival rates than white varieties at low temperatures.

Pea genotypes were Screened for studying winter hardiness at -9C which eliminated all the spring varieties. Romack ID 89-1 and C8M23 had the intermediate hardiness characteristics. A winter hardy field pea variety Austrian Winter, with a compact rosette habit survived at -13C (P. Singh, 1986). Selection at a low temperature and acclimatization under low temperatures are required to develop a cold tolerant variety.

Frost tolerance: Potato is susceptible to frost but a number of genotypes showing resistance to frost have been reported. Frost resistance in potato is controlled by four independent genes. Screening for frost tolerance in potato is possible by the temperature induced chlorophyll fluorescence method (sundbon, E. and Halligren, J.B., 1982).

A high degree of resistance was shown by Solanum depexum, S. aemulens, and S .punea but the species differ in capacity for regeneration. S. phujera & S. stenototum had high regeneration ability. But in S. rybinii, only about a 25% regeneration ability was present (M.A. Vavilova and S.A. Anikina, 1980). Frost resistant species had higher content of easily extractable forms of chlorophyll and carotenoids as compared to non hardy species.

The backcross method was used to develop a frost resistant variety. In the cross of S. tuberosum and S. megistacrolobum, back cross to S. tuberosum produced the line which is resistant to frost and phytophthora. In sweet potato, sugar and dry matter content of the tubers was positively associated the degree of low temperature tolerance. In tomato, L. hirstum f. glabratum and L. pimpinellifolium survived against frost.

Salt tolerance: The degree to which growth and normal metabolism of plant can be maintained in saline soils is described as salt tolerance. Increasing salinity of the soil has become a serious problem in arid and semi arid regions and thus limits the production of vegetable crops. Improvement of salt tolerance in the cultivated species is economical to grow crops under saline conditions.

In L. esculentum screening studies have not results any satisfactory degree of tolerance. Therefore wild species were included in the screening program. In L. peruvianum and L. esculentum minor there was no reduction in fruit size but L. esculentum show considerable reduction in fruit size under saline conditions (Tal.,M. 1971). According to Rush and Epstein, the biotypes of L. cheesmani can survive in full strength sea water in a hydroponic medium, where as L. esculentum cannot sustain even half strength. Tolerance was associated with the ability of the cells to cope with increased sodium levels.

Salt tolerance in Lycopersicon is a heritable characteristic (Rush D.W. and Epstein,E., 1981). The segregating population between Lycopersicon cheesmanii, a tolerant and L. esculentum, a susceptible genotype indicated that tolerance is genetically controlled. In cucurbitaceous crops seed germination was reduced with increasing of salinity in the order Luffa> bottle gourd> round gourd > water melon > bitter gourd > musk melon (Mangal and singh, 1985)

Abelmoschus esculentus is sensitive to salinity. Six varieties were screened at 4.2, 7.8, 11.6 & 16.3mmhos/cm. Pusa sawani was found to be most tolerant to salinity among other varieties for germination and vegetative growth. This variety can be grown in soil having salinity levels up to 6mmhos/cm with Nacl and 8mmhos/cm NaSO4 without significant lose in yield and quality (Mangal, J.L., 1971).

In Aliium cepa, 30 varieties were screened at 4.0, 7.5, and 11.0 mmhos/cm. Salt concentrations >4mmhos/cm reduces the germination. Varieties Hisar-2, Punjab selection, and Udaipur 102 were found to be most tolerant to salinity. Pea varieties were screened at 4, 6, 8, and 12mmhos/cm. In Pea varieties, New line perfection, Market prize, and Duke of Albany were found to be salinity tolerant and can be grown under moderate saline conditions.

Salt tolerance induction in some vegetable crops

Name of crop Variety Name of chemical/ growth substance Cycocel Method of Time for Concentrat treatment treatment ion (hr) Reference

Cauliflower Okra Onion Potato

Hisar-1

250ppm

Root dipping Seed soaking Root dipping

Singh & Mangal (1982) Mangal et al, (1988) Malik, (1973) Yadav et al, (1991) Mangal et al, (1988)

Pusa sawani Hisar-2 Kufri badshah Sel-7

Cycocel Cycocel Sodium salt or cycocel Sodium salts

500ppm 1%

6 8 2

6 ds/m EC Tuber or 250 ppm soaking 8 ds/m EC Root dipping

Tomato

Herbicidal tolerance: Vegetables are susceptible to herbicides applied at pre and post emergence stages. There is a varietal differences in certain vegetable crops. 15 varieties of tomato were treated with metribuzin, a member of triazine at the true three leaf stage. The varieties Vision H 1, vision and fire ball were found to be tolerant. Tolerance to metribuzin in tomato increases with seedling age. 'Heinz 1706 was susceptible at 2 weeks age but tolerant at 4 weeks seedling stage (Dorairaj etal., 1983).

In soybean, the variety hood was found to be tolerant to metribuzin whereas coker 102 was susceptible. Triazine tolerance was studied in C. sativus, the progeny of PI 390244 showed tolerance to the application of triazine ( Gray, D., 1980).
o

The Brassica napus x B.oleracea hybrid showed resistance to atrazine (Horn, R.S., 1980).

Problems related to breeding for abiotic stress

Complex traits controlled by several genes These are highly influenced by environmental factors Poorly understood mechanisms and pathways Varieties do not show wider adaptability Creation of controlled atmosphere is problem Poor heritability because of gene interactions Cross signalling between biotic and abiotic factors

Thank U