Analysis of diallel cross for some physical-quality traits in peanut (Arachis hypogaea L.)

The combining abilities for physical-quality traits in peanut were examined to understand the type of gene action governing these traits, and to identify peanut genotypes suitable for use as parents in breeding for quality improvement. The F₁ hybrids including reciprocals from a six-parent diallel cross along with the parents were evaluated in a randomised complete block design. Data were recorded on five quality traits in peanut viz., shelling outturn, 100-pod weight, 100-seed weight, Count and proportion of sound mature seeds. Substantial genetic variability was observed among the hybrids for the traits studied. Diallel analysis indicated that expression of majority of the quality traits is regulated predominantly by additive gene action suggesting possibility of early-generation selection, while non-additive gene action also plays an equally important role in the control of seed size. Significant reciprocal effect for all the traits denoted role of maternal parent in the expression of quality traits and importance of parental selection in quality breeding. Genotypes ICGV 86564 and TPG 41 were good combiners for seed size, while J 11 was a good combiner for improvement of shelling outturn and proportion of mature seeds. Association between general combining ability (GCA) effects and mean performance suggested that the performance per se of the genotype should be a good indicator of its ability to transmit the desirable quality attributes to its progenies. Though performance of crosses was found to be independent of parental GCA status, it is evident that at least one of the parents used in hybridisation should have large pods and seeds for obtaining better segregants.     

The probable genome donors to Arachis hypogaea L. based on arachin seed storage protein

Summary Interrelationships among six diploid species (A. chacoense, A. villosulicarpa, A. batizocoi, A. correntina, A. duranensis and A. cardenasii), tetraploid wild groundnut A. monticola and four accessions of A. hypogaea were studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of total seed proteins and arachin immunoprecipitates. Arachin in A. monticola and A. hypogaea cv. Trombay Groundnut 9 (TG-9) was composed of four acidic subunits (47.5 kd, 45.1 kd, 42.6 kd and 41.2 kd) and one major basic subunit (21.4 kd). The arachin subunit pattern in cv. Spanish Improved (SP) and TG-1 was almost similar to the pattern observed in A. monticola with the exception that the SP lacked the 41.2 kd and TG-1 lacked the 42.6 kd acidic subunits of A. monticola. Seed extracts of all diploid species studied reacted with the anti-arachin antibodies raised against SP arachin. The electrophoretic analysis of immunoprecipitates of diploid species showed a range of acidic subunits from 46.2 kd to 41.2 kd and one or two basic subunits of 21.4 kd and 20.2 kd. None of the diploid species showed the 47.5 kd subunit found in A. monticola or A. hypogaea. Of the diploid species, A. duranensis and A. cardenasii demonstrated two acidic subunits of 45.1 kd and 41.2 kd and a basic subunit of 21.4 kd as found in A. monticola. Likewise A. batizocoi showed two acidic subunits of 45.1 kd and 42.6 kd and the basic subunit of 21.4 kd as was observed in A. monticola. Based on electrophoretic data, our research supports the earlier conclusion that the probable genome donors to A. monticola are A. batizocoi and A. duranensis or A. cardenasii.     

Variation in progenies of an Arachis hypogaea x diploid wild species hybrid

Summary Derivatives of a cross between cultivated peanuts, Arachis hypogaea L. (2n=40), and the wild species collection GKP 10017 (2n=20) were compared morphologically, for leafspot resistance and for yield. The objective of the study was to determine the effects of wild species germplasm on the A. hypogaea genome. The sterile F₁ hybrid which resulted from crossing the two species was treated with colchicine to restore fertility at the 6x ploidy level. The resulting hexaploid was cytologically unstable and progeny lost chromosomes until stability was regained at the 2n=40 chromosome level. Forty-seven characters were used to analyze the variation among plants in the tetraploid interspecific hybrid population. The plants were compared to four cultivated lines plus GKP 10017. Many hybrids were intermediate to the two parents in morphology. Individual traits such as growth habit, pod and seed size, elongation of the constricted area between pods, nodulation and leaflet size were altered by the presence of GKP 10017 germplasm in many of the hybrid plants. Cercospora arachidicola Hori and Cercosporidium personatum (Berk. & Curt.) Deighton resistances were evaluated for all plants. Several hybrids had few lesions due to either leafspot pathogen. In addition, 24 largeseeded interspecific hybrid selections were compared to the cultivated variety NC 5 for yield. Five selections were superior to both parents at p=0.01. Morphology, disease resistance and yields appeared to be greatly influenced by the wild species GKP 10017 germplasm in plants of the interspecific hybrid population. The potentials of using wild species for improvement of the cultivated peanut are discussed.Paper number 5948 of the journal series of the North Carolina Agricultural Research Service, Raleigh, NC 27650. The investigation was supported in part by ICRISAT and SEA-CR grant no. 701-15-51.     

Identification and inheritance of male sterility in groundnut (Arachis hypogaea L.)

Summary Some plants without pods but with gynophores were observed in two F₄ progenies of two crosses of goundnut (Arachis hypogaea L.). The flowers on these plants had translucent white anthers with no or a few sterile pollen grains. Three such plants in the succeeding generation were hand pollinated with pollen from a short-duration Indian cv. JL 24. The resulting F₁ hybrid plants (male sterile x JL 24) were normal. Chi-square tests for segregation for male fertile and male sterile plants in F₂ and F₃ generations indicated that the male sterility in these crosses of groundnut is governed by two recessive genes. We designate these genes as ms₁ and ms₂ with ms₁ms₁ms₂ms₂ being a male sterile genotype.Submitted as ICRISAT J. A. No. 1812.     

Cytology and leafspot resistance in Arachis hypogaea x wild species hybrids

Summary Introgression of germplasm from diploid wild Arachis species to A. hypogaea has great potential for improving pest resistance in cultivated peanuts. This investigation evaluated methods for incorporating exotic germplasm into cultivated peanuts, especially for Cercospora arachidicola Hori resistance. Interspecific hybrids between A. hypogaea (cvs. NC 2 and NC 5) and the wild species A. cardenasii Krap. et Greg. nom. nud. and A. chacoense Krap. et Greg. nom. nud. were analyzed cytologically and for leafspot resistance. All F₁ hybrids were sterile, had irregular meiosis, and very few multivalents. They were highly resistant to C. arachidicola in field tests and had a 10-fold reduction of conidia per lesion in the greenhouse as compared to A. hypogaea cultivars. After colchicine treatments of F₁ hybrids, hexaploids (2n=60) and aneuploids (2n=54, 56, 63) were observed. The hexaploids had up to 18 univalents per pollen mother cell and very few multivalents, indicating a low frequency of intergenomic chromosome pairing. For C. arachidicola resistance, significant differences were not found among wild species parents, F₁ hybrids and two generations of hexaploids. Most hexaploids were stable at 2n=60 and embryos aborted when backcrosses with the respective wild species were attempted. However, when hexaploids were backcrossed to A. hypogaea, several fertile pentaploid (2n=50) offspring were obtained. Use of self-pollinating pentaploids is believed to be the quickest method to recover 40-chromosome hybrid derivatives in these hybrids.     

Inheritance of albinism in certain interspecific and intersubspecific crosses in groundnut (Arachis hypogaea L.)

Summary A study of the several interspecific and inter-subspecific crosses indicated that albinism in groundnut is recessive and in these crosses is controlled by triplicate recessive loci.ICRISAT Paper No. JA-336     

Inheritance of two components of early maturity in groundnut (Arachis hypogaea L.)

Summary Knowledge of inheritance of early maturity or its components is important to groundnut breeders in developing short-duration cultivars. This study was conducted to determine the inheritance of two components of early maturity: days to first flower from sowing, and days to accumulation of 25 flowers from the appearance of first flower, using three groundnut genotypes. Two early-maturing (Chico and Gangapuri) and one late-maturing (M 13) genotypes were crossed in all possible combinations, including reciprocals. The parents, F₁, F₂, F₃, and backcross populations were evaluated for days to first flower from sowing, and for days to accumulation of 25 flowers. The data suggest that days to first flower in the crosses studied is governed by a single gene with additive gene action. Chico and Gangapuri possess the same allele for this component of earliness. Three independent genes with complete dominance at each locus appear to control the days to accumulation of 25 flowers. In crosses between late (M 13) and early (Chico or Gangapuri) parents, a segregation pattern suggesting dominant-recessive epistasis (13 late:3 early) was observed for this component. Segregation in the F₂ generation (1 late:15 early) of both early parents (Chico x Gangapuri) indicated that the genes for early accumulation of flowers in these two parents are at different loci.Submitted as ICRISAT J.A. No. 1557.     

Identification of polymorphic DNA markers in cultivated peanut (Arachis hypogaea L.)

The detection of DNA polymorphism in cultivated peanut (Arachis hypogaea L.) is reported here for the first time. The DNA amplification fingerprinting (DAF) and amplified fragment length polymorphism (AFLP) approaches were tested for their potential to detect genetic variation in peanut. The AFLP approach was more efficient as 43% of the primer combinations detected polymorphic DNA markers in contrast to 3% with the DAF approach. However, the number of polymorphic bands identified using primers selected in both approaches was comparable. In the DAF study, when 559 primers of varying types were screened, 17 (mostly 10-mer types) detected polymorphism producing an average of 3.7 polymorphic bands per primer with a total of 63 polymorphic markers. In the AFLP study, when 64 primer combinations (three selective nucleotides) corresponding to restriction enzymes Eco RI and Mse I were screened, 28 detected polymorphism. On an average, 6.7% of bands obtained from these 28 primer pairs were polymorphic resulting in a total of 111 AFLP markers. Our results demonstrate that both AFLP and DAF approaches can be employed to generate DNA markers in peanut and thus have potential in the marker-assisted genetic improvement and germplasm evaluation of this economically important crop. This revised version was published online in July 2006 with corrections to the Cover Date.     

The role of mutations in intraspecific differentiation of groundnut (Arachis hypogaea L.)

Summary Based on morphological diversity, cultivated groundnut (Arachis hypogaea L.) is classified into two subspecies (fastigiata and hypogaea) and further into four botanical types (Spanish bunch, Valencia, Virginia bunch and Virginia runner). In a cross between two Spanish cultivars belonging to ssp. fastigiata, a true breeding variant (Dharwad early runner) sharing some characters of both the subspecies was isolated. The variant, on mutagenesis with ethyl methane sulphonate (EMS) yielded a very high frequency of mutants resembling all four botanical types. Some of the mutants produced germinal reversions to Dharwad early runner in later generations indicating genetic instability. While most of the revertants bred true, some of the mutants continued to segregate, wherein each botanical group of mutants produced all other botanical types. A detailed analysis of the breeding behaviour of mutants revealed several unusual features (such as homozygous mutations, mutation outbursts, segregation distortions, somatic mutations and multiple character mutations) that could not be explained through conventional mutation theory. In the light of these findings, the role of mutations in evolutionary differentiation of the crop and the probable mode of their origin have been discussed.     

Heterosis in relation to genetic divergence and specific combining ability in groundnut (Arachis hypogaea L.)

Summary The frequency and magnitude of heterosis were examined in relation to genetic divergence among parents in two diallel cross experiments in groundnut. The parents were grouped into clusters based on their diver-gence. The range, mean and standard deviation of the intra-and inter-cluster divergence were used to define four divergence classes. The frequency of heterotic crosses and the magnitude of heterosis for yield and its components were found to be higher in crosses between the parents in intermediate divergence classes than extreme ones. The results agreed well with the overall status of the specific combining ability of these crosses.     

Reciprocal cross differences in a 6×6 diallel set of groundnut (Arachis hypogaea L.) in the F1 generation

Summary Reciprocal cross differences were studied in a 6×6 diallel full set comprising of thirty hybrid combinations of groundnut in the F₁ generation.Reciprocal cross differences were observed for growth habit in four pairs of crosses, for leaf colour, flower colour and stem pigmentation in two pairs of crosses each. It was observed that the inheritance of flower colour, stem pigmentation and testa colour which exhibited different shades of purple colour was likely to be governed by pleiotropic gene(s). Among the quantitative characters significantly positive reciprocal effects were observed in different crosses for number of mature pods per plant, weight of pods per plant and shelling percent. Marked reciprocal cross differences were observed for pod and kernel characters like pod filling, pod beak, pod constriction and testa colour.     

Genetics of nonnodulation in groundnut (Arachis hypogaea L.). Studies with single and mixed Rhizobium strains

Summary Genetic studies of nonnodulation in groundnut were carried out in a cross, NC 17×PI 259747, with a single Rhizobium strain, NC 92, and a native Rhizobium population.The normal nodulation of the parents, F₁ generations and backcross progenies, and the F₂ segregation for nodulation and nonnodulation confirmed that nonnodulation in groundnut is controlled by two duplicate recessive genes.Approved ICRISAT Journal Article No. 211.     

Components of resistance to late leaf spot and rust among interspecific derivatives and their significance in a foliar disease resistance breeding in groundnut (Arachis hypogaea L.)

Late leaf spot (LLS) and rust cause substantial yield losses and reduce the fodder and seed quality in groundnut (Arachis hypogaea L.). Adoption of resistant cultivars by the semi-arid tropic farmers is the best option to overcome yield losses. Knowledge on components of resistance to these diseases should facilitate the development of groundnut cultivars with enhanced resistance to LLS and rust. The objectives of the experiments were to study the genetic variability and relationships among components of resistance to LLS and rust, and assess their significance in disease resistance breeding. Fifteen interspecific derivatives for LLS and 14 for rust and a susceptible control, TMV 2, were evaluated in a randomised complete block design with two or three replications under greenhouse conditions. The experiments were repeated twice. Genotypic differences were highly significant for all the traits studied. Resistance to LLS is due to longer incubation and latent periods, lesser lesions per leaf, smaller lesion diameter, lower sporulation index, and lesser leaf area damage and disease score. Selection based on components of resistance to LLS may not lead to plants with higher retained green leaf area. The remaining green leaf area on the plant should, therefore, be the major selection criteria for resistance to LLS in breeding programs. Resistance to rust is due to longer incubation and latent periods, fewer pustules per leaf, smaller pustule diameter, lower sporulation index, and lesser leaf area damage and disease score. Rust resistant components appear to work additively, therefore, selection based on resistance components together with green leaf area retained on the plant should be the basis of selecting for resistance to rust in breeding programs. ICGV 99005, 99003, 99012, and 99015 for rust and ICGV 99006, 99013, 99004, 99003, and 99001 for LLS are the better parents for use in resistance breeding programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

花生双受精过程及其经历时间

通过石蜡切片技术研究了花生双受精作用的全过程, 结果表明, 开花后1 h花生的花粉管内的生殖细胞进入有丝分裂, 大约3 h完成。开花后8 h左右精子已经释放到胚囊, 开花后9~13 h精核分别与卵细胞和极核或次生核融合。卵细胞受精之后, 直到开花后30 h合子中的2个核仁渐渐融合, 进入分裂期, 合子多数是横分裂, 少部分纵分裂, 开花后32 h合子分裂形成二细胞原胚。合子休眠期为19 h, 极核在受精之前一部分融合成次生核, 极核或次生核受精之后, 在开花后13 h几乎大部分融合结束, 其中的一个极核或次生核中出现雄性核仁, 形成初生胚乳核, 初生胚乳核不经休眠或极短时间的休眠在开花后15 h分裂。本研究提供了花生受精过程的雌雄性细胞融合形态变化与相应经历的时间。确定了花生合子休眠期, 丰富了花生胚胎学资料, 对花生育种、转基因操作等具有重要意义。     

Variation in the grass pea (Lathyrus sativus L.) and wild species

Summary Forty-nine accessions of Lathyrus sativus were studied for morphological variation. Data were analysed using Principal Components Analysis and Cluster Analysis. The variation in 14 species of Section Lathyrus was also evaluated in order to ascertain affinities between L. sativus and other species. L. sativus is a highly variable species, and there is a clear distinction between the blue-flowered forms from south-west Asia, Ethiopia and the Indian subcontinent, and the white and white and blue flowered forms with white seeds which have a more westerly distribution. Differences in vegetative parts may be due to selection for forage types. L. sativus appears to be closely related to L. cicera and L. gorgoni, and this relationship needs further investigation.     

Determination of genetic variation and taxonomy in lentil (Lens Miller) species by chloroplast DNA polymorphism

Summary Chloroplast DNA restriction fragment lenght polymorphisms (RFLP) were used to examine the taxonomic relationships of cultivated and wild lentil (Lens Miller) species and identify the extent of genetic variation in this genus. Twelve accessions representing all Lens subspecies were digested with four hexanucleotide-recognizing restriction endonucleases. These digests randomly surveyed 540 base pairs, or 0.4% of the approximately 125 kilobase lentil chloroplast genome. A high degree of gragment length conservation was seen among members of crossability group I, i.e., L. c. ssp. culinaris, L. c. ssp. orientalis and L. c. ssp. odemensis. Accessions of the two subspecies comprising crossability group II, i.e., L. n. ssp. nigricans and L. n. ssp. ervoides, showed the greatest amount of variation when compared to the cultivated lentil, L. c. ssp. culinaris. Limited variation was observed within subspecies except for L. n. ssp. nigricans, where accessions of the normal cytotype were highly polymorphic to those of the differentiated cytotype. Chloroplast DNA RFLPs reaffirm hypotheses that propose L. c. ssp. orientalis as the progenitor to the cultivated lentil. The implications of this study on taxonomy and genetic resources is also discussed.     

Evaluation of wild Arachis species for abiotic stress tolerance: I. Thermal stress and leaf water relations

The Indian groundnut cultivars have a narrow genetic base. Hence, it was of interest to investigate the genetic variability among wild Arachis species and their accessions for tolerance to thermal stress. A wide variation was observed in leaf morphological characters such as colour, shape, hairiness, length and width and thickness (SLA). The temperature and time required for 50% leaf injury was worked out with limited number of genotypes and was found to be 54°C for 50 min. Among 36 genotypes (having SLA in the range of 66 and 161 cm² g⁻¹) screened, the inherent potential for cold as well as heat tolerance in terms of relative leaf injury (RI) was observed. Thus, based on RI-values, A. glabrata 11824 and A. paraguariensis 12042 were identified as heat-tolerant and cold-tolerant genotypes, respectively while A. appresipila 11786 was found to be susceptible to both heat and cold. Correlation between SLA and RI values for heat (r = 0.38, P < 0.05) and cold (r = 0.52, P < 0.05) tolerance was positive, indicating that thicker the leaf the lower the injury or higher the tolerance. Among six species and 13 accessions, comprising both heat-tolerant and heat- susceptible genotypes, the concentrations of various leaf chemical constituents such as total protein, phenols, sugars, reducing sugar, amino acids, proline, epicuticular wax load and chlorophyll varied significantly. The epicuticular wax load ranged between 1.1 and 2.5 mg dm² among 13 A.glabrata accessions. These accessions were categorized into two groups, i.e. high-wax (range: 2.0–2.5 mg dm²) and low-wax types (range: 1.1–1.6 mg dm²). The high-wax type showed a higher diffusion resistance (dr) as compared to low-wax type; though the transpiration rate (tr) in high-wax type was moderate (between 9.5 and 11.6 μg cm⁻² s⁻¹). Genetic variability in parameters such as canopy temperature, dr and tr was also distinct. The fully turgid leaves with relative water content ≥91%, showed leaf water potential (ψ_leaf) between −0.7 and −1.2 MPa. Results indicated that the plants with thicker leaves are better protected from heat injuries. Further, epicuticular wax load seems to help in maintaining stomatal regulation and leaf water relations, thus affording adaptation to wild Arachis species to thrive under water-limited environments. The sources of tolerance, as identified in this study, could be utilized to improve thermal tolerance of the groundnut cultivars by intra-specific hybridization, following either conventional breeding using embryo rescue techniques, if required or utilizing biotechnological tools.     

花生种间杂种胚胎发育及内源激素变化

以花生栽培品种豫花15为母本分别与野生种A. correntina和A. macedoi杂交,以栽培品种间杂交组合豫花15×白沙1016为对照,采用石蜡切片技术观察花生杂种胚胎发育进程,酶联免疫吸附分析法(ELISA)测定花生杂种胚胎内源激素含量, 分析胚胎发育进程与胚胎内源激素含量的关系。结果显示,3个杂交组合的下针率无明显差异,表明栽培种与2个野生种杂交不存在受精前障碍;与对照相比,豫花15×A. correntina表现杂交可亲和,其胚胎发育中激素含量的变化与对照组合基本一致;而豫花15×A. macedoi杂交授粉后12 d内胚胎发育基本正常,17~27 d发育缓慢,之后珠被迅速向内增生抑制了胚的进一步发育并最终败育,在同一时期的杂种胚胎中,3-吲哚乙酸(IAA)、赤霉素(GA₃)和玉米素核苷+二氢玉米素核苷(ZR+DHZR)等促进生长的激素维持在较低的水平,而脱落酸(ABA)含量以及脱落酸与某一促进生长激素或3类促进生长激素之和的比值保持了较高的水平,推测可能是激素代谢的失衡导致种间杂种胚胎的败育。用IAA、激动素(KT)单独或混合处理不亲和组合豫花15×A. macedoi杂交授粉后的果针基部表明,外施促进生长的激素能够明显增加胚珠的长度,一定程度上验证了上述推测。     

RAPD markers linked to a rust resistance gene in cultivated groundnut (Arachis hypogaea L.)

Groundnut rust (Puccinia arachidis Speg.) is an important air borne pathogen, which causes substantial losses in groundnut yield and quality. Although large numbers of accessions were identified as rust resistant in wild, interspecific derivative and cultivated groundnut species, transfer of resistance to well-adapted cultivars is limited due to linkage drag, which worsens yield potential and market acceptance. A F₂ mapping population comprising 117 individuals was developed from a cross between the rust resistant parent VG 9514 and rust susceptible parent TAG 24. Rust resistance was governed by single dominant gene in this cross. We identified 11 (out of 160) RAPD primers that exhibited polymorphism between these two parents. Using a modified bulk segregant analysis, primer J7 (5′CCTCTCGACA3′) produced a single coupling phase marker (J7₁₃₅₀) and a repulsion phase marker (J7₁₃₀₀) linked to rust resistance. Screening of the entire F₂ population using primer J7 revealed that the coupling phase marker J7₁₃₅₀ was linked with the rust resistance gene at a distance of 18.5 cM. On the other hand, the repulsion phase marker J7₁₃₀₀ was completely linked with rust resistance. Additionally, both J7₁₃₀₀ (P = 0.00075) and J7₁₃₅₀ (P < 0.00001) were significantly associated with the rust resistance. Marker J7₁₃₀₀ identified all homozygous rust resistant genotypes in the F₂ population and was present in all the eight susceptible genotypes tested for validation. Thus, J7₁₃₀₀ should be applicable for marker-assisted selection (MAS) in the groundnut rust resistance breeding programme in India. To the best of our knowledge this is the first report on the identification of RAPD markers linked to rust resistance in groundnut.     

Intra species variation in Atylosia scarabaeoides (L.) Benth., a wild relative of pigeonpea (Cajanus cajan (L.) Millsp.)

Summary Atylosia scarabaeoides (L.) Benth., a wild relative of pigeonpea, possesses several useful genes which can be utilized for pigeonpea improvement. In the present study, 33 accessions of A. scarabaeoides were evaluated at ICRISAT Center during the 1987 rainy season for variation in some useful traits to identify parents for inter-generic hybridization. A large variation was observed for leaf components, seed size, pod length, seeds/pod, days to flowering, seed protein, sulphur amino acids, resistance to cyst nematode, phytophthora blight, sterility mosaic, fusarium wilt, pod borer, pod fly, and pod wasp. Only four accessions were found to have more than 28% protein content. Methionine and cystine contents were marginally higher than in pigeonpea but the variation was not large enough to utilize them in the breeding program. In A. scarabaeoides. accessions resistant to fusarium wilt, phytophthora blight, sterility mosaic, and cyst nematode were detected. Compared to pigeonpea, the A. scarabaeoides accessions were less susceptible to lepidopteran borer and were immune to pod fly damage. Accessions ICPW 89 and ICPW 111 in short- (100–120 days), and ICPW 94 and ICPW 118 in medium-duration (140–180 days) were identified as potential parents for use in inter-generic hybridization.ICRISAT Journal Article No. 967