NUTRITIONAL IMPROVEMENT IN VEGETABLE CROPS

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NUTRITIONAL IMPROVEMENT IN VEGETABLE CROPS Amit & Koundinya

APPEAL:

APPEAL AN APPEAL IS MADE BY THE AUTHORS TO GO EASY ON THEM IF ANY COPYRIGHT VIOLATION IS FOUND IN THIS PRESENTATION

NUTRITIONAL STATUS:

NUTRITIONAL STATUS 53% of children of the world are malnourished and underweight and 40% of them live in India 70% of malnourished people live in South & Southeast Asia. Nearly 800 million people remain hungry & nearly 1.3 billion live in less than $1US / day Nearly 19000 infants among 40000 die every day due to malnutrition related disorders. 3.2 billion live with iron & vit A deficiency resulting irreversible blindness among 125 million school children. The Hindu (2005) Bagchi and Hazra (2005)

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WHO, Geneva

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Diseases

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NUTRIENTS VEGETABLES Carbohydrate sweet potato, potato, garlic, lima bean Protein pea, lima bean, cowpea Fat Chilli, sweet pepper, brinjal, brussel’s sprout Vitamin A Carrot, palak, spinach, amaranthus Vitamin B 1 Tomato, chilli, garlic, leek, pea Vitamin B 2 Palak , chilli , sweet pepper, cole crops VEGETABLES AGAINST MALNUTRITION

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NUTRIENTS VEGETABLES Vitamin C Chilli, sweet pepper, cabbage, drumstick Calcium Hyacinth, bean, amaranthuns, palak Iron Amaranthus, palak, spinach, lettuce Phosphorous Garlic, pea, lima bean, taro, drumstick leaves Vitamin B 5 Palak, amaranthus, bitter gourd, pointed gourd Iodine Tomato, sweet pepper, carrot, garlic Sodium Celery, green onion, Chinese cabbage, radish

PHYTOCHEMICALS (ANTIOXIDENT NUTRIENTS) :

PHYTOCHEMICALS (ANTIOXIDENT NUTRIENTS) Organic compounds from plants Health promoting properties Low in fat, no cholesterol Found in small amount in vegetables Protect from chronic diseases

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ANTIOXIDANT VEGETABLES Beta- carotene carrots, orange bell peppers, pumpkins, and sweet potatoes winter squash, spinach, Vitamin-C Broccoli, Brussels sprout, green leafy vegetables cabbage, green pepper, tomato Lutein / zeaxanthin Red pepper, Kale, broccoli, spinach, winter squash, Brussels sprout, celery, leak, mustard green, pea, green onion, summer squash Major antioxidants supplying vegetables (Cont.)

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Lycopene,tarpnoid Tomato and water melon Vitamin-E Green leafy vegetables and sweet potato drum stick fenugreek Lipoic acid Green leafy vegetables, especially spinach, beat and broccoli Flavonoids Bitter gourd (Lee et al 2004) Anthocynanin Blue Swede var of potato Phenolic compounds Pepper, egg plant (Cao.et.al 1996) ANTOXIDANT VEGETABLES

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Antioxident content in different vegetables

TOXICANTS OF VEGETABLES:

VEGETABLES TOXIC COMPOUNDS ADVERSE EFFECTS Carrot Carotatoxin (polyacetylenic alcohol Neurotoxic symptoms Lettuce Nitrates , alkaloids Mathemoglobinemia Brassica (cruciferous vegetables) Gluosinolates, choline-esterase inhibitor, S-methyl cysteine sulfoxides Goiter, digestive disorders Beets, spinach Oxalates, nitrates, phytate, tannins, saponins, nitrosamine Mathemoglobinemia , reduces bioavailability of certain minerals such as calcium, iron, zinc. Carcinogenic TOXICANTS OF VEGETABLES (cont.)

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VEGETABLES TOXIC COMPOUNDS ADVERSE EFFECTS Sweet potato Ipomeamqarone Enzyme inhibitors Water melons Serotonin Pumpkin and squashes Choline-esterase inhibitor Neurotoxic Legumes (vegetables) Lectins, cyanogenic glycosides, hemagglutinins, trypsin, amylase, glucose-6-p-dehydrogennase inhibitor, compounds having anti vitamin properties (vit-A, E and D) Allergens Asparagus Sponins , choline -esterase inhibitor Neurotoxic (cont.)

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VEGETABLES TOXIC SUBSTANCES ADVERSE EFFECTS Potato Solanine Chaconine Invertase inhibitor Burning sensation Tomato Tomatine Gastric discomfort Pungent pepper (chillies) capsaicin Skin irritation, gastric disorders Lettuce, Carrot, Onion Tannins Epithelium and Liver damage Cassava CN glucosides Legume vegetables Protease inhibitors Pancreatic hypertrophy Leafy vegetables (Amaranths, Spinach) Oxalic compounds Kidney stones Crucifer vegetables Goitrogens Anti thyroid effects Kubota et al., 2010

HOW TO IMPROVE NUTRIENT CONTENT?:

HOW TO IMPROVE NUTRIENT CONTENT? By the Application of Plant Nutrients By Plant Breeding

NUTRIENT IMPROVEMENT BY THE APPLICATION OF PLANT NUTRIENTS:

NUTRIENT IMPROVEMENT BY THE APPLICATION OF PLANT NUTRIENTS CROP NUTRIENTS IMPROVES REFRENCES Capsicum DAP+CAN Vitamin C Basavaraja et al. (1998) K 2 O@130kg/ha Vitamin C Bubiez et al. (1981) Chilli K 2 O@60kg/ha Capsaicin, Ascorbic acid Ananthi et al. (2004) Tomato K 2 O@100kg/ha Protein content Pansare et al.(1994) Onion P 2 O 5 @80 kg/ha Amino acid Balasubramanian et al.(1978) Water melon Etheral@250ppm TSS Dixit et al. (1999) GA 3 @25ppm TSS

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July 2006, UAS Dharwad

BREEDING APPROACHES:

BREEDING APPROACHES Evaluation of Germplasm Hybridization Somaclonal Variation Mutagenesis Interspecific Hybridisation Genetic Engineering

OBJECTIVES & PRIORITIES OF Nutritional breeding:

OBJECTIVES & PRIORITIES OF Nutritional breeding Consumer’s acceptance Yield cannot be scarified for nutritional qualities Identification & selection against compounds with toxic & /or antinutritional compounds. Emphasis on nutritional quality for maximum impact on human nutrition & wellness. Exploitation of genetic variability. Genetic manipulation for biosynthesis of chemical compounds. (Fredrick, 1999)

EVALUATION OF GERMPLASM:

EVALUATION OF GERMPLASM It consists of screening of germplasm including varieties for nutritional traits along with good agronomic features. European and Asiatic carrot were evaluated for carotene, dry matter and TSS content (Gill and Kataria,1974) In tomato 42 genotypes were evaluated in 3 replications at 2 locations. Pant Bahar,CLN-31-0-4-2,CLN2123A,CLN2123E, CLN2143B, CLN1621L, AC1017, AC1037, AC897 are promising for TSS % over a wide range of atmosphere. (Aradhana Joshi,2003)

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Even though there was no significant difference among the cultivars andersen had the highest TSS content. This could be used in further breeding programmes

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Crop Traits present Varieties References Tomato Vit.C TSS Lycopene & β –carotene Pusa Red Plum ( 33mg/ 100 g) Double Rich, Redrock Hisar Arun Red Cherry, Novelty Angurlata , Pusa Ruby Caro Red, High Pigment Thamburaj , 1998 Rai & Rai , 2006 Thomes,1988 Kalloo , 1988 Harihar Ram, 2000 CULTIVATED VARIETY

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Crop Traits present Varieties References Pea Protein GC 195, kinnauri , Laxton , Swaroop , 2006 Pumpkin Carotene Golden Delicious Neamtu,1982 Carrot Vit - A TSS Pusa Meghali Nantes,Surkh chanteny Maliva,1984 Pepper Carotene Douxed Alger Thamburaj , 1998 Musk melon TSS Arka Rajahans , Durgapur madhu Swaroop , 2006 Water melon TSS Arka Muthu 12-14% IIHR, 2010

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Cassava / tapioca- Starch 28% H226, Sree Prakash Sree Sahya, Swaroop, 2006 Industrial starch Sree Harsha, Co.2 Starch 35.6% MVD 1 Potato Dry matter Protein &vit.C Starch K.Dewa, K.Red K.Chandramukhi K.Sindhuri Kufri chipsona 1,2,3&4 Hawkes,1990 Swaroop , 2006

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CROP VARIETY TRAIT Tomato Double rich, Hisar Arun High Vit C Chilli/ Paprika NIC 19953 Color extraction EC 347263 capsaicin Muskmelon PSDPM 1-7-50 High TSS Carrot HCO-1 Beta Carotine Onion Phule safed, white onion, No. 569 High TSS Rai & Rai, 2006

GERMPLASM LINE:

GERMPLASM LINE Crop Traits Accession no. References Tomato TSS Ascorbic acid Citric acid A276, A408, UC828 EC 5425 PI 263713 Rai & Rai , 2006 Rana et al., 1982 Stavens , 1986 Pea Protein K 1554,AC141, P23, EC21857 Gupta et al ., 1984 Potato Vitamin C Protein Dry matter JEX/A Nos. 63,68, 208, 218, 226, 276, 426, 493 JEX/A Nos. 68, 275, 403, 592 QB/A-9-120 King & Birhman , 1993

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Crop Traits Accession no. References Capsicum spp. Carotene Lines WA,SY, HP, AB Sharma et al., 1989 Carrot Vitamin A Carotene EC178385 EC277678, EC187207 Gautam et al., 1998

Wild Relatives:

Wild Relatives Wild relatives of a crop includes its primitive forms (from where they have originated), related weedy species and other species in the same genus, which are not under cultivation These may serve as potential sources of genes for nutritional improvement of cultivated forms

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Crop Wild relative Traits References Tomato L.hirsutum L.pimpinellifolium β carotene Vit.C , TSS Stommel et al. 1994 L. hirsutum & glabratum L. cheemanii L. minutum Carotene TSS Thamburaj 1998 L. peruvianum Vit.C Clutter, 1986 Brinjal S. khasianum S. aviculare Low solasodine Bose et al., 2003 Capsicum C. annuum var aviculare Capsiacin Chadha et al., 1988

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Crop Wild relative Traits References Cucumber C. Sativus var xishaung bannansis Carotene Qi et al., 1983 Potato S.microdontum S.vernei S.phureja spp. phureja Calcium Starch Carotene John Bambey , 2003

HYBRIDIZATION:

HYBRIDIZATION Hybridization includes crossing between two genetically diverse genotypes. In some cases (polygenic nutritional characters) selection can also be followed after hybridization- Pedigree method of breeding. Monogenic nutritional characters can be brought into cultivated types by following back cross method of breeding. Recurrent selection is followed to increase desirable gene frequencies and to break undesirable linkages. RS for 2 cycles increased seed protein in French bean from 21.9% to 24.6% (Sullivan,1981) RS and RRS for increasing TSS content in tomato (Kalloo, 1988)

HETEROSIS BREEDING FOR NUTRITIONAL TRAITS:

HETEROSIS BREEDING FOR NUTRITIONAL TRAITS CROP HYBRID ATTRIBUTES REFRENCES Tomato Arka Vikas x Sel 12 Ascorbic acid Bhatt et al , 1998 Bhutani et al , 1980 KS10 x Pant T 3 Pusa red plum X Punjab Chhuhara Best of All X HS101 PT 10 x Pant Bahar TSS and Ascorbic acid Kumar et al, 2001 Pant Bahar X Pusa Ruby G-2 x HS 110 Carotenoid and Lycopene Bhutani et al , 1980 L. esculentum cv Rutgers x L. hirsutum Carotene Tomes et al ,1978 (Cont.)

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CROP HYBRID ATTRIBUTES REFRENCES Cabbage Sel -1 X Sel- 36 Asocrbic acid Peter, 1998 Musk melon ms 2 X Pusa madhuras TSS 13.5% TSS 26% Munshi et al. 2005 Arka Jeet x Sarda melon Water melon Mateera AHW19 x Sugarbaby Vitamin A High TSS Samadia et al ., 2002 Peter, 1998 Crimson sweet X New hampshire Chilli Punjab guchnedar X LLS Capsaicin Oleoresin High vit C Rai & Rai, 2006 PKM-1 x Arka Lohit CH-3 Spinach beet IIHR P-6 x Pusa Harit Protein Singh and Sainii , 1998

Inheritance of Nutritional Traits:

Inheritance of Nutritional Traits TRAITS CROP GENETICS Carbohydrate / TSS Pea Recessive gene Water melon, Muskmelon, Tomato Polygenic Carrot Single major gene (Rs) Protein Pea Add. & dom. gene Soybean Dominant gene Broad bean Polygene & modifiers Vitamin A Tomato, Chilli Add. Gene Carrot Complentary gene Cucurbita sp. Polygenic

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TRAITS CROP GENETICS Vitamin C Cucumis melo L., Chilli Add. & dom. gene Tomato Additive gene Pea Polygenic Dry Matter Tomato Polygenic (3 genes) with additive effects Capsaicin Chilli Monogenic Carotenoid Carrot Complex Inheritance Kalloo, 1988

INTERSPECIFIC HYBRIDISATION:

INTERSPECIFIC HYBRIDISATION High quality lines derived from such crosses usually serve as parents in hybridisation program. Lycopersicon esculentum x Lycopersicon hirsutum – high beta carotene Lycopersicon esculentum x Lycopersicon hirsutum f glabratum – high beta carotene ( Kalloo, 1988 )

MUTAGENESIS:

MUTAGENESIS Spontaneous mutation Induced mutation Mutants can be serve as source of quality genes Sweet Potato Mutant “Port Blaco” from Port Rico - White flesh, High starch ( Thamburaj,1998 )

MUTANTS :

MUTANTS Crop mutagen Remarks References Tomato γ rays Spontaneous High dry matter & sugar Increased β -carotene Hvostova et al ., 1976 Konsler et al., 1973 Capsicum γ rays EMS Irradiation High capsaicin content 20-60% capsaicin increment High protein Sigurbjournsson et al ., 1974 Abdalla, 1983 Gotts chalk, 1983 Sweet potato EI Variation in sugar content Kukimura et al., 1975 B. oleracea Spontaneous Orange curd mutant Cripset et al ., 1975

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Irradiation-induced mutation breeding is effective in improving sweet potato characters such as yield, starch and soluble sugar content, carotenoids content of storage roots (Wang et al., 2007) Shin et al., (2011) evaluated gamma radiated Sweet potato genotypes and found higher starch and protein content in mutagenic material than normal ones

SOMACLONAL VARIATIONS (SCV):

SOMACLONAL VARIATIONS (SCV) The generation of considerable variation, which is heritable during tissue culture is known as SCV (Rai & Rai 2006).

SOMACLONAL VARIATION:

SOMACLONAL VARIATION Genetic variation present in tissue cultured raised plants “Scarlet” variety of sweet potato developed through soma clonal variation is having high starch content. Tomato variety “ DNAP-9”, 20% more soluble solids (Liu, 2010)

SOMACLONAL VARIANTS:

SOMACLONAL VARIANTS Crop Somaclonal variants observed Tomato (DNAP 9) High solids in fruit Carrot High carotene content Sweet potato High starch content (Liu, 2010)

Transgenics…..:

Transgenics ….. A plant in which a foreign has been transferred through genetic engineering is called a transgenic plant and the gene so transferred is called transgene. (Singh, B.D. 2001)

TRANSGENES IN VEGETABLES:

TRANSGENES IN VEGETABLES CROP TRAIT GENE RESPONSIBLE GENE SOURCES Tomato ↑ Sucrose, ↓ Starch Sucrose phosphate synthase Maize Potato ↑ Starch(20-40%) ADP-glucose pyrophosrylase(ADP-GPPase) E.Coli ↑ Starch Starch synthase _ ↓ Sugar Invertase inhibitor( inhh ) Tobacco Water melon ↑ TSS ADP-glucose pyrophosphate Sweet Potato (Berry et al , 1999)

Engineering for Nutrition:

Engineering for Nutrition Grain Amaranthus - A. hypochondriacus contain higher storage grain protein content. This protein contain 2-4 folds more essential amino acids – Methionine, Lysine, Leucine & Threonine than normal ones The gene responsible for this protein AmA1 has been transferred successfully into Potato Patented by scientists at NBPGR, New Delhi, headed by Dr. Asis Dutta. (Chakraborty et al. 2000 JNU, ND)

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88 AmA1 transgenic lines of 9 Indian potato cultivars viz. Kufri Badshah, Kufri Bahar, Kufri Chipsona-1, Kufri Chipsona-2, Kufri Jyoti, Kufri Lauvkar, Kufri Pukhraj, Kufri Sindhuri and Kufri Sutlej were developed at CPRI, Shimla. Expression of AmA 1confirmed in tubers of 48 transgenic lines. Evaluated twice at Modipuram and Shimla each. Data on protein quality and quality evaluated.

FOR STARCH ENHANCEMENT:

Potato var. with high sugar content leads to charring of sugar while chips making Potato varieties with high starch content are needed by Food industry. ADPGPP- ADP Glucose Pyro Phosphorylase, the first enzyme in the pathway of starch biosynthesis . This gene has been transferred to potato cultivars from E.coli, which resulted in dramatic increase in starch content in potato tubers (Monsanto, USA). FOR STARCH ENHANCEMENT

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Sugar (Glucose, Fructose) GBSS Alpha Amylase gene Antisense promoter Starch Alpha Amylase m-RNA Antisense Alpha Amylase gene m- RNA m- RNA Inhibits Starch conversion into sugar STARCH ENHANCEMENT (Kuipers et al . 1994)

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In plants acid invertase is thought to be involved primarily in conversion of sucrose to glucose & fructose for energy in growing tissues and increasing extracellular invertase activity results in severely stunted growth and inhibition of photosynthesis (Schaewen et al., 1990). In tomato invertase action is inhibited by using antisense gene technology thereby inhibited the sucrose metabolism

Grafting for nutritional improvement:

Grafting for nutritional improvement Grafting is one of the useful tool for improving yield and nutritional quality Flavor, pH, sugar, color, carotenoid content, and texture can be affected by grafting and the type of rootstock used (Davis et al. 2008) An increase in ascorbic acid content in tomato was found with grafting (Zhu et al . 2006)

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Savvas et al ., 2011 Effect of different rootstocks on TSS and Ascorbic acid content in tomato cv. Belladona

ACHIEVEMENTS AT NATIONAL LEVELS:

ACHIEVEMENTS AT NATIONAL LEVELS Crop Varieties Developed by Attributes References Potato K. Chipsona 1 K.Chipsona 2 CP2416xMS78-79 F-6xQBIB92-4 High dry matter, low sugar Paul et al ., 2002 Water melon Mateera AHW 19 Mateera AHW 25 Selection from local race  TSS Pareekh et al ., 2002 Cow pea Bidhan Barbati 1 Bidhan Barbati 2 Combination involving 3 sub species  Protein % Hazra et al ., 2001

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Crop Varieties Developed by Attributes Reference Spinach Arka Anupama IIHR10x IIHR 8 High Vit.A, C, Carotene & Fe Varalakshmi et al., 2001 Amaranth Arka Arunima Pure line selection from IIHR 49 Rich in Ca, Fe, low antinutional factors Varalakshmi et al., 2001 Musk melon MH4-5 Durgapur Madhu x Hara Madhu  TSS (13-16%) Lathura et al., 2002 Water melon RW-177-3 (Durgapur Lal) Sugar Baby x K-3566  TSS (10-11%) Yadav et al., 2002

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ACHIVEMENTS AT INTERATIONAL LEVEL CROP VARIETY DEVELOPED BY ATTRIBUTES REFERANCES Potato Amflora, Fortuna BASF Ltd. High industrial starch content Anonymous, 2011 Eliane AVEBE Ltd. 100% amylo pectin Anonymous, 2011 Tomato T4063,T4099, T5019,T5020 Inbreeding and backcrosses High level of carotenoids Wann et al ., 1997 USDA 97L63,97L66, 99L97 L.esculantum cv.Floradade, L.cheesmani ( LA317) High beta carotene Stommel , 2001

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CROP VARIETY DEVELOPED BY ATTRIBUTES REFERANCES Cucumber EOM400, EOM402,LOM404 Triple cross High carotene Jack et al., 2011 Carrot B7262 PI173687 x B10138 High carotene(100-130mg/kg) Simon et al ., 1997 Beta Sweet Selective breeding >40% beta carotene Sterling, 2003 Sweet potato NPSP-01,02,03,04,05,06 Open pollination of 24 parents High dry matter Mwanga , 2003

LIMITATIONS :

LIMITATIONS Polygenic nature of some nutritional traits like protein content. Negative correlation between yield & nutritional quality. Transfer of character from wild spp. may pose several problems such as cross incompatibility, hybrid inviability, hybrid sterility & linkage of undesirable genes with desirable ones. Such work involves lot of chemical analysis. The analysis requires close cooperation of biochemist which sometimes becomes limiting factor in the progress. Low heritability of some nutritional traits.

FUTURE THRUST:

FUTURE THRUST Restrengthening the germplasm collection. Development of F1 hybrids, varieties and induce mutants with higher nutritional values. Exploitation of molecular biology and cellular genetics for development of transgenics with high nutritional traits. Human nutritionists must specify the objectives a breeder is expected to achieve in breeding for nutritional improvement. Incorporation of nutritional traits should be integrated with the normal breeding procedures for yield. A close intraction between nutritionists and breeder will increase the rate of progress in breeding for nutritional quality.

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