Exploitation of wild annual Cicer species for widening the gene pool of chickpea cultivars

Introgression of unadapted genes from the wild Cicer species could contribute to the widening of genetic base of important traits such as yield, yield attributes and resistance to major biotic and abiotic stresses. An attempt was made successfully to intercross two wild annual Cicer species with three cultivated chickpea cultivars. Four interspecific cross‐combinations were made, and their true hybridity was ascertained through morphological and molecular markers. These cross‐combinations were also studied for some important quantitative traits under real field conditions. The range, mean and coefficient of variation of agro‐morphological traits were assessed in the parental lines, their F₁ and F₂ generations to determine the extent of variability generated in cultivated chickpea varieties. A high level of heterosis was recorded for number of pods/plant and seed yield/plant in F₁ generation. Three cross‐combinations of ‘Pusa 1103’ × ILWC 46, ‘Pusa 256’ × ILWC 46 and ‘Pusa 256’ × ILWC 239 exhibited substantially higher variability for important yield‐related traits. The present research findings indicate that these wild annual Cicer species can be easily exploited to broaden the genetic base of cultivated gene pool for improving seed yield as well as adaptation.     

Transgressive segregations for yield criteria in reciprocal interspecific crosses between <Emphasis Type="Italic">Cicer arietinum</Emphasis> L. and <Emphasis Type="Italic">C. reticulatum</Emphasis> Ladiz.

Wild Cicer species are considered to be more resistant for biotic and abiotic stresses than that of the cultivated chickpea (Cicer arietinum L.). Alien genes conferring resistance for biotic and abiotic stresses can be transferred from wild Cicer species to the cultivated chickpea but success in interspecific hybridizations has already been achieved with only two wild Cicer species. The current study was undertaken to compare fruitful heterosis in F₂ and F₃ for yield and yield criteria and to identify transgressive segregation in F₂ and F₃ in reciprocal interspecific crosses between C. arietinum and C. reticulatum Ladiz. We define fruitful heterosis as a useful parameter that can be used instead of residual heterosis. Considerable fruitful heterosis in F₂ and F₃ was found for number of seeds, pods per plant, biological yield, and seed yield. Maximum values of most of the characteristics in F₂ and F₃ were higher than that of the best parent indicating that superior progeny could be selected for yield from transgressive segregation. Progeny selection should be based on number of seeds, pods per plant, and biological yield since these characteristics had the highest direct effect on seed yield. The narrow sense heritability was found to be the highest for 100-seed weight. It was suggested that the cultivated chickpea could be used as female parent in interspecific hybridization to increase yield and yield criteria since progeny in F₂ and F₃ had better performance when it was used as female. In conclusion, interspecific hybridization of wild and cultivated chickpea can be used to improve yield and yield components and resistance to biotic and abiotic stresses as well.     

Cytogenetic study of Helianthus laevigatus and its F1 and BC1F1 hybrids with cultivated sunflower, Helianthus annuus

Although the wild sunflower species Helianthus laevigatus has not been extensively studied it may be considered for sunflower breeding as a potential source of desirable genes for Sclerotinia stalk rot resistance and high contents of proteins and linoleic acid in the seed. A set of six H. laevigatus populations was crossed to cultivated sun~ower lines and produced nine F₁ (2-14 plants) and 66 BC₁F₁ hybrid combinations (1-13 plants). Male sterility occurred in F₁ and BC₁F₁ hybrid combinations and pollen viability was lower in the progenies than in the parents (51.6-77.2%in F₁ and in F₁ and 4.8-34.0% in BC₁F₁). Meiosis was normal in the H. laevigatus populations It was found that this tetraploid species also occurred in a hexaploid form Numerous irregularities were observed in the meiosis of the F₁ interspecific hybrids During diakinesis, quadrivalents and hexavalents were recorded in addition to bivalents Dislocated chromosomes and chromosome bridges were present in the other phases The chromosome number in F₁ was 68 (tetraploid). Irregularities in chromosome pairing were observed in the interspecific hybrids at BC₁F₁. There were many univalents, and trivalents quadrivalents and hexavalents were also present The chromosome number in the BC₁F₁ generation ranged from 34 to 60. The occurrence of meiotic irregularities in the F₁ and BC₁F₁ interspecific hybrids indicates that H. laevigatus and the cultivated sunflower differ in genome constitution.     

Reproductive behaviour and broomrape resistance in interspecific hybrids of sunflower

Interspecific hybrids and backcross generations between the wild perennial species Helianthus resinosus, Helianthus paucifiorus, Helianthus laevigatus, Helianthus nuttallii ssp. nuttallii T. & G. and Helianthus giganteus, resistant to broomrape (Orobanche cernua) and susceptible inbred lines were obtained to study crossability to cultivated sunflower and the transmission and expression of resistance to this parasitic weed. Conventional crosses with all the species tested were successful except for the crosses with diploid H. giganteus, for which embryo rescue techniques were needed to overcome hybrid incompatibility. Pollen viability and seed set were highest for F₁ hybrids with hexaploid species and lowest for those with the diploid H. giganteus. We evaluated F₁, BC₁F₁, some BC₂F₁ plants and the wild and cultivated parents. The wild species and interspecific hybrids were resistant to broomrape infection except for H. nuttallii, which showed segregation, indicating that the resistance is dominant. The crossability and resistance of F₁, and back-cross generations of species with different ploidy levels indicate that the transfer of broomrape resistance to cultivated sunflower is feasible.     

Development of Bruchid-Resistant Mungbean Line Using Wild Mungbean Germplasm in Thailand

A mungbean (V. radiata) line (BC₃F₃ generation) which is resistant to two species of bruchid beetles (Callosobruchus chinensis and C. maculatus) was successfully developed in Thailand using a wild mungbean variety (V. radiata var. sublobata). One accession (TC1966) of wild mungbean was found to be completely resistant to C. chinensis and C. maculatus occurring at Chainat Field Crops Research Center in Thailand. The resistance was controlled by a single dominant gene (R). A breeding program to develop a bruchid-resistant mungbean cultivar with good agronomic characters under the environmental conditions of Thailand was initiated in 1987.‘Chainat 60’ (‘CN60’), a recommended mungbean cultivar in Thailand, was crossed with TC1966 to incorporate the resistance gene. Agronomic characters of the hybrids were improved by recurrent backcrossing using ‘CN60’ as a pollen parent. Seed yield per plant, days to flowering, and seed size of the bruchid-resistant BC₃F₂ population reached the level of ‘CN60’ after three consecutive backcrossings. Bruchid-resistant line (BC₃F₃, R/R) was selected from individual BC₃F₂ plants.     

Introgression of productivity and other desirable traits from ricebean (Vigna umbellata) into black gram (Vigna mungo)

Crosses were performed to introgress genes for productivity and other desirable traits from ricebean (Vigna umbellata) into black gram (Vigna mungo). Crossability was very poor in black gram × ricebean crosses, and only two to nine true hybrid plants were obtained. Plant fertility was very poor in initial generations, but was improved gradually from F₂ onwards. Twenty‐four uniform progenies, bulked in F₇, were evaluated for yield potential. The percentage increase/decrease in yield ranged from ?35.48 to 50.31 over the check cultivar (‘Mash338’, female parent). All the progenies were found resistant to Mungbean yellow mosaic virus, Cercospora leaf spot and Bacterial leaf spot diseases. Overall, it was found that desirable traits such as high pod number, seed weight, productivity and resistance to diseases have been introgressed successfully into black gram from ricebean. A derivative line, KUG114, recorded yield superiority of 39.45% over the check cultivar ‘Mash338’ on the average of 14 multilocation research trials. It was released under the name ‘Mash114’ for cultivation in the Punjab state.     

Inheritance of seed protein in two interspecific pigeonpea (Cajanus cajan) crosses

Pigeonpea is one of the primary sources of protein for most rural communities of south Asia and Africa. The enhancement of protein from the present level of 20–22% will help masses in their nutritional well-beings. Breeders have identified high-protein (28–30%) germplasm amongst the wild relatives of pigeonpea; and these can serve as donors in breeding high-yielding high-protein cultivars. To achieve this, information on the inheritance of seed protein will be useful in organizing interspecific matings, generation advancement, and selection schemes. In the present study, two interspecific crosses, involving two pigeonpea cultivars ('Pant A2' and 'ICP 6997') and a wild species (Cajanus albicans (W. & A.)), were made to study the inheritance of seed protein in F₁, F₂, and BC₁F₁ generations. The high-protein trait was found to be controlled by two independent dominant genes with complementary effects. These genes are tagged with symbols “HPt₁” and “HPt₂”. The authors believe that now opportunities exist for enhancing the protein levels in pigeonpea through breeding approaches.     

Introduction of aromatic fragrance into cultivated tomato from the peruvianum complex'

This study was performed to introduce the distinct aromatic fragrance of Lycopersicon peruvianum LA 1554 into the cultivated tomato, Lycopersicon esculentum. The strong breeding barriers existing between these two distantly related species were circumvented by the ovule selection and culture method. A large BC₁F₁ population was developed and among 127 plants, 36 were self‐compatible and yielded fruits. Fruits of some of these selected plants were found to be enriched with a sweet aromatic flavour. Sensory evaluation of the fruit aroma of these selected plants was performed by a panel of 12 members against one of the best consumer‐rated Japanese commercial tomato cultivars, ‘Momotaro’. Although extensive variation was observed in fruit‐aroma in the BC₁F₁ population, panel opinion on ‘flavour‐desirability’ significantly favoured the BC₁F₁ fruits of some selected plants over the cv. ‘Momotaro’. Therefore, it can be concluded that the aromatic fragrance of a ‘L. peruvianum’ accession has successfully been introduced into the cultivated tomato gene pool.     

Results in the Development of Alloplasmic Carrots (Daucus carota sativus Hoffm.)

Alloplasmic forms of the cultivated carrot, Daucus carota sativus Hoffm., were created by substitution backcrosses. The wild Daucus forms D. muricatus L., D. gingidium L., D. c. gummifer Hook, and D. c. libanotifolia Wiinst. were used as cytoplasm donors. In the first two backcross generations disturbances in fertility and plant physiology did not occur. The plants of the generations BC₂F₂ and BC₂F₃ look like cultivated carrots. The alloplasmic forms were compared with cultivars for economic traits such as yield and yield components. The high content of total sugar in the alloplasmic forms was considerable. A new type of male sterility was found and described in a BC₃F₂ generation of the cross D. c. gummifer× D. c. sativus. Results of RFLP-analyses of mt-DNA and cp-DNA were discussed.     

Introgression of resistance to Columbia and Northern root-knot nematodes fromSolanum bulbocastanum into cultivated potato

Summary Resistance toMeliodogyne chitwoodi races 1 (MC1) and 2 (MC2) andM. hapla (MH) derived fromSolanum bulbocastanum was introduced into the cultivated potato gene pool through somatic fusion. The initial F₁ hybrids showed resistance to the three nematodes. Resistance to reproduction on roots by MC1 was accompanied by resistance to tuber damage in F₁ clones. Tuber damage sometimes occurred, however, in hybrids of BC₁ progeny resistant to reproduction on roots when MC2 and MH were the challenging nematodes. Resistance to reproduction was transferred into BC₁ individuals, but a greater proportion of BC₁ progeny was resistant to MC1 than to MC2 or MH. Resistance to MC1 appears to be dominant and discretely inherited. F₁ and BC₁ progeny were pollen sterile, but seed were produced from crosses using cultivated tetraploid pollen sources. Approximately 11 and 33 per cent of pollinations produced berries on F₁ and BC₁ pistillate parents, respectively. Seed yield increased fourfold overall in crosses with F₁ compared to BC₁ individuals.Abbreviations MC1 Meloidogyne chitwoodi race 1 - MC2 Meloidogyne chitwoodi race 2 - MH Meloidogyne hapla - Rf Reproductive factor     

Introgression of Cajanus platycarpus genome into cultivated pigeonpea, C. cajan

Summary Cajanus platycarpus, an incompatible wild species from the tertiary gene pool of pigeonpea (C. cajan (L.) Millspaugh), has many desirable characteristics for the improvement of cultivated varieties. To necessitate such transfers, embryo rescue techniques were used to obtain F₁ hybrids. The F₁ hybrids were treated with colchicine to obtain tetraploid hybrids, that were selfed to obtain F₂, F₃ and F₄ progenies. All of the hybrids and subsequent progenies had an intermediate morphology between the two parents. Backcrossing of the tetraploid hybrids with cultivated pigeonpea was not possible given embryo abortion, with smaller aborted embryos than those obtained in the F₀ parental cross.As a route of introgression, diploid F₁ hybrids were backcrossed with cultivated pigeonpea and BC₁ progeny obtained by in vitro culture of aborting embryos. BC₂ plants were obtained by normal, mature seed germination. Although embryo rescue techniques had to be used to obtain F₁ and BC₁ plants, it was possible to produce BC₂ and subsequent generations through direct mature seed. Every backcross to cultivated pigeonpea increased pollen fertility and the formation of mature seeds.Special project assistant till December, 2003.     

Development of high yielding IR64 × <Emphasis Type="Italic">Oryza rufipogon</Emphasis> (Griff.) introgression lines and identification of introgressed alien chromosome segments using SSR markers

Modern rice varieties that ushered in the green revolution brought about dramatic increase in rice production worldwide but at the cost of genetic diversity at the farmers’ fields. The wild species germplasm can be used for broadening the genetic base and improving productivity. Mining of alleles at productivity QTL from related wild species under simultaneous backcrossing and evaluation, accompanied by molecular marker analysis has emerged as an effective plant breeding strategy for utilization of wild species germplasm. In the present study, a limited backcross strategy was used to introgress QTL associated with yield and yield components from Oryza rufipogon (acc. IRGC 105491) to cultivated rice, O. sativa cv IR64. A set of 12 BC₂F₆ progenies, selected from among more than 100 BC₂F₅ progenies were evaluated for yield and yield components. For plant height, days to 50% flowering and tillers/plant, the introgression lines did not show any significant change compared to the recurrent parent IR64. For yield, 9 of the 12 introgression lines showed significantly higher yield (19–38%) than the recurrent parent IR64. Four of these lines originating from a common lineage showed higher yield due to increase in grain weight and another three also from a common lineage showed yield increase due to increase in grain number per panicle. For analyzing the introgression at molecular level all the 12 lines were analyzed for 259 polymorphic SSR markers. Of the total 259 SSR markers analyzed, only 18 (7.0%) showed introgression from O. rufipogon for chromosomes 1, 2, 3, 5, 6 and 11. Graphical genotypes have been prepared for each line and association between the introgression regions and the traits that increased yield is reported. Based on marker trait association it appears that some of the QTL are stable across the environments and genetic backgrounds and can be exploited universally.     

Diallel analyses reveal the genetic control of resistance to ascochyta blight in diverse chickpea and wild Cicer species

Ascochyta blight is a major fungal disease affecting chickpea production worldwide. The genetics of ascochyta blight resistance was studied in five 5 × 5 half-diallel cross sets involving seven genotypes of chickpea (ICC 3996, Almaz, Lasseter, Kaniva, 24B-Isoline, IG 9337 and Kimberley Large), three accessions of Cicer reticulatum (ILWC 118, ILWC 139 and ILWC 184) and one accession of C. echinospermum (ILWC 181) under field conditions. Both F₁ and F₂ generations were used in the diallel analysis. The disease was rated in the field using a 1–9 scale. Almaz, ICC 3996 and ILWC 118 were the most resistant (rated 3–4) and all other genotypes were susceptible (rated 6–9) to ascochyta blight. Estimates of genetic parameters, following Hayman’s method, showed significant additive and dominant gene actions. The analysis also revealed the involvement of both major and minor genes. Susceptibility was dominant over resistance to ascochyta blight. The recessive alleles were concentrated in the two resistant chickpea parents ICC 3996 and Almaz, and one C. reticulatum genotype ILWC 118. The wild Cicer accessions may have different major or minor resistant genes compared to the cultivated chickpea. High narrow-sense heritability (ranging from 82% to 86% for F₁ generations, and 43% to 63% for F₂ generations) indicates that additive gene effects were more important than non-additive gene effects in the inheritance of the trait and greater genetic gain can be achieved in the breeding of resistant chickpea cultivars by using carefully selected parental genotypes.     

Mapping a major gene for growth habit and QTLs for ascochyta blight resistance and flowering time in a population between chickpea and <Emphasis Type="Italic">Cicer reticulatum</Emphasis>

Ascochyta blight is a devastating disease of chickpea. Breeders have been trying to introduce resistance from wild Cicer into cultivated chickpea, however, the effort is hampered by the frequent genetic drag of undesirable traits. Therefore, this study was aimed to identify potential markers linked to plant growth habit, ascochyta blight resistance and days to flowering for marker-assisted breeding. An interspecific F₂ population between chickpea and C. reticulatum was constructed to develop a genetic linkage map. F₂ plants were cloned through stem cuttings for replicated assessment of ascochyta blight resistance. A closely linked marker (TA34) on linkage group (LG) 3 was identified for plant growth habit explaining 95.2% of the variation. Three quantitative trait loci (QTLs) explaining approximately 49% of the phenotypic variation were found for ascochyta blight resistance on LG 3 and LG 4. Flowering time was controlled by two QTLs on LG3 explaining 90.2% of the variation. Ascochyta blight resistance was negatively correlated with flowering time (r = −0.22, P < 0.001) but not correlated with plant growth habit.     

Influence of Phytohemagglutinin on the Agronomic Performance of Beans (Phaseolus vulgaris L.)

The abundant lectin phytohemagglutinin (10 % of total seed protein) does not contain sulfur amino acids and, being a potent antimetabolite, it is responsible for the lowering of the nutritional value of bean seeds. The aim of the present work was to improve the dry bean cultivar ‘Taylor's Horticultural’ (Asgrow), by genetically introducing the lectin null (lec/lec) character from two null genotypes: ‘Pinto UI 111’ and ‘Heidi’. Thirty-seven BC²F₃ and fourteen BC₆F₅ inbred lines were evaluated in agronomical trials. Analysis of Variance (ANOVA) showed significant differences among BC₂F₃ breedings lines for all traits under evaluation. Comparison of the LedLee genotypes versus lec/lec did not show statistically significant differences in the means for the following traits: yield, yield components and percentage of protein in the seed. Fourteen BC6F5 lines, compared together with their recurrent parent ‘Taylor's Horticultural’, showed significant differences among genotypes for 1000 seed weight, protein percentage on dry matter and ash percentage. No significant differences were observed for grain yield. The data indicate that lectin removal did not have a detrimental effect on the traits evaluated.     

Broadening the genetic base of lentil cultivars through inter‐sub‐specific and interspecific crosses of Lens taxa

Wild Lens taxa are invaluable sources of useful traits for broadening genetic base of cultivated lentil. Nine inter‐sub‐specific and interspecific crosses were made successfully between cultivated (Lens culinaris ssp. culinaris) and wild lentils (L. culinaris ssp. orientalis, odemensis, lamottei and ervoides). The effect of species groups, day length and temperature on crossability in lentils was evident under normal winter sowing in New Delhi and in summer Himalayan nursery at Sangla in Himachal Pradesh, India, although pollen fertility assessed in all the cross‐combinations showed no significant variation. True hybridity of nine inter‐sub‐specific and interspecific crosses was confirmed through morphological and molecular (ISSR) markers, in which three of 120 primers could confirm the hybridity of all the crosses. All cross‐combinations were also studied for important quantitative traits related to yield. The range, mean and coefficient of variation were estimated in parental lines, F₁ and F₂ generations to determine the extent of variability generated in cultivated lentils through the introgression of genes from wild L. taxa. A high level of heterosis was observed in F₁ crosses for important traits studied. Substantially higher variations for seed yield and its attributing traits were exhibited in F₂ generations indicating transgressive segregation. The results of the present investigation revealed that wild L. taxa can be successfully exploited for lentil improvement programmes, and the variations generated could be easily utilized for broadening the genetic base of cultivated lentil gene pool for improving the yield as well as wider adaptation.     

Interspecific and intergeneric hybridization and chromosomal engineering of Brassicaceae crops

In Brassicaceae crop breeding programs, wild relatives have been evaluated as genetic resources to develop new cultivars with biotic and abiotic stress resistance. This has become necessary because of the diversification of ecotypes of diseases and pests, changing food preferences, advances in production technology, the use of new approaches such as in vitro breeding programs, and the need for economical production of F₁ seed. To produce potential new cultivars, interspecific and intergeneric hybridizations have been performed between cultivated species and between cultivated species and their wild relatives. Furthermore, interspecific and intergeneric hybrids have been successfully produced using embryo rescue techniques. In this paper, we review the interspecific and intergeneric incompatibilities between Brassicaceae crops and their wild relatives, and the production, characterization, and improvement of synthetic amphidiploid lines, alien gene introgression lines, alloplasmic lines, monosomic alien chromosome addition lines, and monosomic alien chromosome substitution lines. The goal is to provide useful materials to support practical breeding strategies and to study the genetic effects of individual chromosomes on plant traits, the number of genes that control a trait, their linkage relationships, and genetic improvement in Brassicaceae crops.     

Molecular marker-assisted selection for development of common bean lines resistant to angular leaf spot

The objective of this work was to develop homozygous common bean lines carrying angular leaf spot resistance genes derived from the cultivars ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’ through marker‐assisted selection. Molecular markers SCAR OPN02₈₉₀, RAPD OPE04₅₀₀ and OPAO12₉₅₀ linked to the resistance genes of ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’, respectively, were used in segregating backcross‐derived populations to selection. DNA fingerprinting was used to select homozygous BC₂F₃ and BC₁F₃ resistant plants genetically closer to the recurrent parent. Two homozygous BC₂F_2:3 and two and five BC₁F_2:3 families derived from ‘Ruda’ vs. ‘Mexico 54’ (RM), ‘MAR 2’ (RMA) and ‘BAT 332’ (RB) crosses were selected, respectively. After only one (RMA, RB) or two backcrosses (RM), five and eight BC₁F₃ lines derived from RMA and RB, respectively, and seven BC₂F₃ lines derived from RM, with 14.9–16.6, 16.9–18.6 and 9.3–11.1% of relative genetic distances to the recurrent parent were selected. This is the first report of lines resistant to angular leaf spot carrying genes of the cultivars ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’ developed with the aid of molecular markers.     

Breeding cowpea for resistance to Striga gesnerioides in the Nigerian dry savannas using marker‐assisted selection

Historically, conventional breeding has been the primary strategy used to develop a number of Striga‐resistant varieties currently grown in the Sahel of Western Africa. In this study, we have successfully developed and applied a marker‐assisted selection strategy that employs a single backcross programme to introgress Striga resistance into farmer preferred varieties of cowpea for the Nigeria savannas. In this strategy, we have introduced the Striga resistance gene from the donor parent IT97K‐499‐35 into an elite farmer preferred cowpea cultivar ‘Borno Brown’. The selected 47 BC₁F₂ populations confirmed the recombinants with desirable progeny having Striga resistance gene(s). The 28 lines selected in the BC₁F_2:4 generation with large seed size, brown seed coat colour and carrying marker alleles were evaluated in the field for resistance to Striga resistance. This led to the selection of a number of desirable improved lines that were immune to Striga having local genetic background with higher yield than those of their parents and standard varieties.     

Transfer of new dwarfing genes from the weed species Avena fatua into cultivated oat A. byzantina

New sources of dwarfing genes were identified from accessions of Avena fatua in Japan and Korea. The dwarfing genes were transferred from backcrossed and self‐pollinated relatives to the cultivated oat ‘Kanota’. In the cultivated form, the dominant dwarfing gene Dw8 showed a relatively lower transmission rate than recessive, semi‐dominant and nondwarfing genes and was characterized by a distinct link with wild gene cluster. This was also supported by the high transmission rate of wild specific SSR alleles. Four dwarf inbred lines (L153, L169a, L169b and L812) were identified as involving a single recessive dwarfing gene(s). The recessive dwarfing genes that showed normal and stable transmission rates in BC₁F₃ were first reported in hexaploid oats. The L169 segregated two different recessive dwarf lines in BC₁, which were selected as semi‐ (L169a) and extreme‐ (L169b) dwarf lines. The L169a was a good genotype with a high grain yield. L288 is a semi‐dwarf line conditioned by a semi‐dominant dwarfing gene, with a unilateral panicle, large florets and good grain quality due to strong resistance to lodging. L342 had a short peduncle, making the panicle compact, and its phenotype was similar to the dwarfness controlled by Dw7, but the dwarfing genes were different.