A mechanism for inhibiting cross-fertilization in pigeonpea (Cajanus cajan (L.) Millsp.)

Summary Natural out-crossing imposes considerable costs and inefficiencies in breeding, evaluation and commercialization of pigeonpea (Cajanus cajan (L.) Millsp.). This note reports identification of a modification of floral morphology which inhibits cross-fertilization. Floral morphology and possible mechanisms of action of this character are discussed.On leave from International Crops Research Institute for the Semi-Arid Tropics (ICRISAT). Hyderabad, India.     

Possibility of genetic improvement of pigeonpea (Cajanus cajan (L.)Millsp.) utilizing wild gene sources

Summary Various wild relatives of pigeonpea,Cajanus cajan, namely some species ofAtylosia andRhynchosia, possess desirable characteristics that could be utilized for effecting genetic improvement of this crop. In total 73 cross combinations among two cultivars ofC. cajan and one accession each of eightAtylosia species and one ofRhynchosia were attempted. Twelve hybrids were obtained. Seven of these were analysed for F₁ fertility and their utility for agronomic improvement of theC. cajan. Fertility behaviour of the different F₁ hybrids varied and indicated that potential of gene transfer between the two genera,Atylosia andCajanus, was as good as within the genusAtylosia. From F₂ and F₃ families ofC. cajan × A. scarabaeoides andC. cajan × A. albicans, plants were selected with greater physiological efficiency and agronomic superiority. The prospects of transferring pod borer resistance and higher seed protein content from someAtylosia species to pigeonpea are discussed.     

A possible gene source for early,day-length neutral pigeon peas,Cajanus cajan (L.) Millsp.

Summary Atylosia scarabaeoides Benth. and A. platycarpa Benth., close relatives of the species Cajanus cajan (L.) Millsp., were screened for photoperiodic response. Four photoperiods ranging from 12 h 45 min to 19 h were studied in three environments. A. scarabaeoides flowered freely only in the first photoperiod. A. platycarpa not only flowered early (39 to 63 days after planting) in all four photoperiods, but also exhibited a relatively constant vegetative phase up to 16 h of illumination. Cajanus cajan, in which most cultivars-if not all-exhibit a response to photoperiod, was successfully hybridized with the two Atylosia species. It is suggested that genes for earliness and insensitivity to day-length could be transferred from A. platycarpa to C. cajan.     

RFLP analysis of cytoplasmic male-sterile lines of pigeonpea [Cajanus cajan (L.) Millsp.] developed by interspecific crosses

Total DNA from three putative cytoplasmic male sterile (CMS) progenies derived from crosses between the wild species Cajanus sericeus and the cultivated species Cajanus cajan, five C. cajan, one accession of C. sericeus and two genetic male sterile lines of pigeonpea were compared for their RFLP patterns using maize mitochondrial DNA (mtDNA) specific probes. Three putative cytoplasmic male sterile (CMS) progenies from the multiple cross genome transfer of pigeonpea lines (CMS 7–1, CMS 12–3, and CMS 33–1) showed hybridization patterns identical to that of C. sericeus when DNA was digested with EcoRI and HindIII and probed with maize mtDNA clones. The results suggested that these putative CMS progenies have the mitochondria of the female wild species parent. The hybridization patterns of the three male parental lines used in the development of the CMS progenies were similar in all the restriction enzyme-probe combinations except HindIII-atp6. The genetic male sterile lines, MS Prabhat and QMS 1 differed from each other in their hybridization pattern. The genomic DNA hybridization pattern of HindIII digested DNA from ICPL 87 differed from the other pigeonpea lines when probed with the maize mtDNA clones. The cluster analysis of the hybridization data suggested the occurrence of variation in the mitochondrial genome even among the cultivated species. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Microsporogenesis and anther wall development in male-sterile and fertile lines of pigeon pea (Cajanus cajan (L.) Millsp.)

Summary Microsporogenesis and anther wall development of male-sterile and fertile lines in pigeon pea (Cajanus cajan (L.) Millsp.) was examined microscopically. Male-sterility was complete and was caused by breakdown at the young tetrad stage. Degeneration of the tapetum by vacuolation occurred during the first division of meiosis and appeared responsible for pollen mother cell breakdown. Sterile plants also differed from the fertile plants in the enlargement of the inner middle layer of the anther wall, and in the lack of development of the endothecium.     

Species affinities between Cajanus cajan and some Atylosia species based on esterase isoenzymes

Summary Esterase isozymes were studied in seed extracts of Cajanus cajan and six Atylosia species by polyacrylamide gel electrophoresis and isoelectrofocusing. The isozyme patterns were stable and accession specific. Within the accessions of the Atylosia species, A. albicans and A. scarabaeoides showed three common bands indicating that they are more closely related to each other than to the other species. Of the accessions of Atylosia only A. cajanifolia shares the esterase isozyme of C. cajan and hence seems to be the closest wild relative of C. cajan.     

Pollen mother cell and anther wall development in a photoperiod-insensitive male-sterile mutant of pigeon pea (Cajanus cajan (L.) Millsp.)

Summary The development of the pollen mother cells (PMC's) and anther wall of a male-sterile mutant was microscopically compared with that of fertile sib plants in pigeon pea (Cajanus cajan). Male-sterility was complete and caused by breakdown of microsporogenesis at prophase I. Delayed and incomplete development of the anther wall appeared to be responsible for PMC degeneration. The sterile mutant also differed from the fertile plants in the size and number of the PMC's.On leave from International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India.     

Variation for natural out-crossing in pigeonpea

Summary Several researchers have reported a considerable degree of natural out-crossing in pigeonpea from different environments. This paper reviews the subject with respect to the variation for natural out-crossability, pollinating vectors, extent of natural out-crossing, isolation specifications, and the possible utilization of natural out-crossing in pigeonpea improvement.ICRISAT Journal Article No. 831.     

Inheritance pattern for pod lenghth and ovule number in wide crosses among Cajanus and Atylosia species

Summary Six interspecific/intergeneric F₂ populations involving two cultivars of C. cajan and five Atylosia species were scored for segregation patterns for pod length and ovule number. It was observed that for both characters the F₂ means and the ranges were correspondingly lower than the midparent values and the spread of the two parents. However, the intrageneric cross of A. lineata and A. scarabaeoides showed transgressive segregation for pod length and the F₂ mean and range were correspondingly larger than the midparent value and spread of the two parents, suggesting the usefulness of combining ability test for choice of suitable parents.     

Inheritance of stem termination in pigeon pea (Cajanus cajan (L.) Mill sp.)

Summary Inheritance studies on the stem termination in pigeon pea using F₁, F₂ and F₃ generations of two crosses between determinate and indeterminate lines suggested that two dominant genes with epistatic (inhibitory) interaction of one of them control the interminate growth habit. The gene symbols D. idid and ddIdId have been designated to the parental plants with determinate and indeterminate growth habits, respectively. The gene IdId was epistatic (inhibitory) to the gene D giving a ratio of 13 indeterminate: 3 determinate inthe F₂'s observed. F₃ segregation supported the proposed model on the mode of inheritance.     

Random amplified polymorphic DNA analyses of cytoplasmic male sterile and male fertile Pigeonpea (Cajanus cajan (L.) Millsp.)

Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of two pigeonpea cytoplasmic genic male sterile (CMS) lines derived from crosses between the wild (Cajanus scarabaeoides & C. sericeus) and the cultivated species of Cajanus cajan. The male sterile (A) line and its maintainer (B) line could be easily differentiated with certain random primers. The two male sterile (288 A and 67 A)systems are based on C. scarabaeoidesand the other is based on C. sericeus could also be differentiated. Amplification product of 600bp amplified by primer OPC-11 was observed in both the cytoplasmic male sterile lines (288 A and 67A), which was absent in the maintainer lines (288 B and 67 B) and the putative R-line (TRR 5 and TRR 6). CMS lines, putative R lines, other cultivars and wild species under the study could be easily distinguished with the help of different primers. Dendrogram constructed based on the similarity index showed that considerable genetic variation exist sbetween CMS lines, two putative R line and wild species studied. This revised version was published online in July 2006 with corrections to the Cover Date.     

Diversity in selected wild and cultivated species of pigeonpea using RFLP of mtDNA

Diversity in 28 accessions representing 12 species of the genus, Cajanus arranged in 6 sections including 5 accessions of the cultivated species, C. cajan, and 4 species of the genus Rhyncosia available in the germplasm collection at ICRISAT was assessed using RFLP with maize mtDNA probes. Cluster analysis of the Southern blot hybridization data with 3 restriction enzymes – 3 probe combinations placed the genus Rhyncosia in a major group well separated from all the species belonging to the genus Cajanus. Within the genus Cajanus, the 4 accessions of C. platycarpus belonging to section Rhynchosoides formed a separate group in contrast to those in other sections of pigeonpea. In the section, Cajanus all the 5 accessions of C. cajan were grouped together and C. cajanifolius belonging to the same section was in a subgroup by itself closer to the main group. The four accessions of C. scarabaeoides, were together and the other species belonging to section Cantharospermum were in different subgroups. The intra-specific variation was seen even within accessions of certain pigeonpea wild species such as C. scarabaeoides, C. platycarpus, C. acutifolius, and even the cultivated species of C. cajan. This study suggests that RFLP of mtDNA can be used for the diversity analysis of pigeonpea and it gives some indications on the maternal lineage among the species. The variations in the mitochondrial DNA hybridization patterns also suggest the extensive rearrangement of the organelle genome among the Cajanus species. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Seed protein profiles of pigeon pea (Cajanus cajan) and some Atylosia species

Summary The seed protein profile of pigeon pea (Cajanus cajan) appeared to be highly stable. The standard profile of this cultigen was found in 86 of the 90 accessions examined. Each of the Atylosia species, A. lineata, A. platycarpa, A. cajanifolia and A. scarabaeoides has its own profile but each band of the Atylosia species had a homologue in the standard profile of Cajanus or in one of its variants. This was taken as an indication for the polyphyletic origin of Cajanus from several Atylosia species. The appearance of specific Atylosia bands in some of the electrophoretic variants of Cajanus suggests that gene flow is still effective between pigeon pea and various Atylosia species. The poor solubility of the Atylosia seed protein in comparison to Cajanus indicates that domestication of Cajanus was coupled with increased solubility and perhaps a better nutritional value of this pulse as well.     

Inheritance of trichomes and resistance to pod borer (Helicoverpa armigera) and their association in interspecific crosses between cultivated pigeonpea (Cajanus cajan) and its wild relative C. scarabaeoides

The legume pod borer, Helicoverpa armigera, is one of the most devastating pests of pigeonpea. High levels of resistance to pod borer have been reported in the wild relative of pigeonpea, Cajanus scarabaeoides. Trichomes (their type, orientation, density and length) and their exudates on pod wall surface play an important role in the ovipositional behavior and host selection process of insect herbivores. They have been widely exploited as an insect defense mechanism in number of crops. In the present investigation, inheritance of resistance to pod borer and different types of trichomes (A, B, C and D) on the pod wall surface in the parents (C. cajan and C. scarabaeoides) and their F₁, F₂, BC₁ (C. cajan × F₁), and F₃ generations has been studied. Trichomes of the wild parents (high density of the non-glandular trichomes C and D, and glandular trichome B and low density of glandular trichome A) were dominant over the trichome features of C. cajan. A single dominant gene as indicated by the segregation patterns individually will govern each trait in the F₂ and backcross generation. Segregation ratio of 3 (resistant): 1 (susceptible) for resistance to pod borer in the F₂ generation under field conditions was corroborated with a ratio of 1:1 in the backcross generation, and the ratio of 1 non-segregating (resistant): 2 segregating (3 resistant: 1 susceptible): 1 non-segregating (susceptible) in F₃ generation. Similar results were obtained for pod borer resistance under no-choice conditions. Resistance to pod borer and trichomes associated with it (low density of type A trichome and high density of type C) are governed individually by a dominant allele of a single gene in C. scarabaeoides. Following backcrossing, these traits can be transferred from C. scarabaeoides into the cultivated background.     

Chloroplast-DNA variation in the wild and cultivated olives (Olea europaea L.) of Morocco

The male sterile plants that segregated in a BC₅F₂ of `C. sericeus × C. cajan var. TT-5' population were maintained by sib mating. The male sterile plants were crossed with ICPL-85012.Approximately 50% of the F₁ plants were sterile. F₂ plants derived from the fertile F₁ plants did not segregate for male sterility. The reciprocal hybrid i.e. ICPL-85012 × Fertile derivatives from C. sericeus × TT-5, did not express male sterility. However, among the 12 F₂ plant to row progenies, two segregated 25% male sterile plants and remaining 10 did not segregate. The segregation pattern in subsequent progenies revealed that the sterility was under control of a single recessive allele. Studies on the backcross and their BC₁F₂ and BC₁F₃progenies revealed another sterility gene which was found to be dominant in inheritance. This paper shows that what was thought to be cytoplasmic male sterility from C. sericeus cytoplasm is actually a single dominant gene possibly acting in concert with a single recessive gene to mimic cytoplasmic male sterility. This revised version was published online in July 2006 with corrections to the Cover Date.     

Natural out-crossing in dwarf pigeonpea

Summary Natural out-crossing rate in pigeonpea was studied at ICRISAT Center using plant stature (tall plants in dwarf progenies) as the genetic marker. The data indicated natural out-crossing rates of 9.7% to 24.1% with a pooled value of 13.1% in the six populations studied. These data were comparable to earlier studies at the same site using stem colour and growth habit as genetic markers in tall pigeonpea cultivars thus suggesting that foraging of insect pollination vectors is not influenced by plant type. The implications of natural out-crossing on breeding and maintenance of genetic purity of cultivars is discussed.     

Role of partial cleistogamy in maintaining genetic purity of pigeonpea

Summary Natural out-crossing in pigeonpea (Cajanus cajan (L.) Millsp.) leads to genetic deterioration and in certain environments the out-crossing may be as high as 70%. Studies with partially cleistogamous lines derived from an inter-generic cross indicated that average natural out-crossing in these lines was much lower (2.5%) than that reported earlier for pigeonpea (36.5%) at ICRISAT Center. Based on seed yield and disease resistance, potential parents such as ICPL 87154 in short-duration determinate, ICPL 87018 in short-duration indeterminate, and ICPL 87159 in medium-duration indeterminate groups have been identified for introgression of the partially cleistogamous trait into pigeonpea.