Phylogenetic relationships of Cajanus cajan (L.) Millsp. (pigeonpea) and its wild relatives based on seed protein profiles

Total seed storage protein of 9 accessions of cultivated C. cajan and 10 wild Cajanus species was reported and compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis. A considerable variation was detected among the protein profiles of different accessions of C. cajan while those of wild species were very specific and distinctly different from each other. Relative similarities between various taxa were estimated by Jaccard's similarity index and cluster analysis was performed to produce a UPGMA dendrogram. The clustering of 10 wild species and C. cajan more or less agrees with their sectional classification and available data based on morphological characteristics, crossability, genome pairing in hybrids and nuclear RFLPs. The species closest to C. cajan is C. cajanifolia although the accessions of C. cajan also share some bands present in the profiles of C. scarabaeoides, C. goensis, C. lineatus, C. acutifolius and C. volubilis. This points towards polyphyletic origin of the cultigen which has been discussed in this paper.     

AFLP Fingerprinting in Pigeonpea (Cajanus cajan (L.) Millsp.) and its Wild Relatives

Detection of DNA polymorphism in cultivated pigeonpea (Cajanus cajan) and two of its wild relatives Cajanus volubilis and Rhynchosia bracteata is reported here for the first time using amplified fragment length polymorphism (AFLP) fingerprinting. For this purpose, two EcoRI (three selective nucleotides) and 14 MseI (three selective nucleotides) primers were used. The two wild species shared only 7.15% bands with the pigeonpea cultivars, whereas 86.71% common bands were seen among cultivars. Similarly, 62.08% bands were polymorphic between C. volubilis and pigeonpea cultivars in comparison to 63.33% polymorphic bands between R. bracteata and pigeonpea cultivars, and 13.28% polymorphic bands among pigeonpea cultivars. The cluster analysis revealed low polymorphism among pigeonpea cultivars and very high polymorphism between cultivated pigeonpea and its wild relatives. The AFLP analysis also indicated that only one primer combination (EcoRI + ACT and MseI + CTG), at the most any four primer pair combinations, are sufficient for obtaining reliable estimation of genetic diversity in closely related cultivars like pigeonpea material analyzed herein. AFLP analysis may prove to be a useful tool for molecular characterization of pigeonpea cultivars and its wild relatives and for possible use in genome mapping.     

Effect of temperature on flavonoid production in pigeonpea [Cajanus cajan (L) Millsp] in relation to nodulation

To ascertain the reasons for poor nodulation of pigeonpea, we studied the effect of high temperature on the production of flavonoids by the pigeonpea host. A high temperature affected flavonoid production by pigeonpea and mungbean. At 37°C pigeonpea root exudates contained four flavonoids and the root extract contained five. The proportion of the second flavonoid in the pigeonpea and the mungbean was higher and the proportion of the third flavonoid was lower at 37°C compared to 30°C. At the higher temperature the flavonoids exuded from pigeonpea roots were same those in the root homogenate.     

Effect of high temperature on plasmid curing of Rhizobium spp. in relation to nodulation of pigeonpea [Cajanus cajan (L.) Millsp.]

Summary Fifty-six isolates of Rhizobium and Bradyrhizobium spp. (Cajanus) were studied for their plasmid profile and N₂-fixation efficacy. One to three plasmids were reproducibly detected in all the Rhizobium spp. strains but no plasmid was detected in the Bradyrhizobium spp. strains. Rhizobium sp. strain P-1 was mutagenized by Tn5 and three nod^– and six nod⁺fix^– were screened for symbiotic parameters. Neomycin-sensitive mutants were isolated by elevated temperatrue (40°C) from tranconjugants carrying Tn5 insertions. The high temperature cured these mutants from the single large plasmid present in the parent strain P-1. All these cured mutants were nod^–, indicating that the genes for nodulation were present on this plasmid, which is readily cured at a high temperature (40°C). The high temperature in the semi-arid zones of Haryana could be responsible for the low nodulation of pigeonpea because the plasmid carrying the nodulation genes is cured at 40°–45°C giving rise to non-nodulating mutants.     

Impact of nodule damage by Rivellia angulata on N2-fixation,growth, and yield of pigeonpea (Cajanus cajan L. Millsp.) grown in a Vertisol

Summary Damage caused by Rivellia angulata larvae to pigeonpea root nodules at the ICRISAT center in India was greater in the crop grown on Vertisols (up to 86%) compared to that on Alfisols (20%). Attempts to quantify the field effects of nodule damage on growth and yield of pigeonpea in a Vertisol, involving many heavy applications of soil insecticides (aldrin and hexachlorocyclohexane) failed because the insecticides did not control the pest and adversely affected the growth of the pigeonpea and the subsequent crop of sorghum (Sorgorum bicolor L. Moench). The impact of nodule damage on pigeonpea growth, yield and nutrient uptake was successfully studied in greenhouse-grown plants at three N levels. In this pot study, artificial inoculation with Rivellia sp. led to substantial nodule damage (70%). The results of this damage were a significant overall reduction in nodule dry weight (46%), acetylene reduction activity (31%), total leaf area (36%), chlorophyll content of leaves (39%) and shoot dry weight (23%) 68 days after sowing. At maturity, Rivellia sp. infestation caused significant reductions in top dry weight (22%), root and nodule dry weight (27%), seed dry weight (14%), and total N (29%) and P uptake (19%). The problems and prospects of manipulating nodule damage so as to reduce N losses in pigeonpea are discussed.Submitted as JA No. 756 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Comparative salinity tolerance of symbiotically dependent and nitrogen-fed pigeonpea (Cajanus cajan) and its wild relative Atylosia platycarpa

Summary Once symbiosis between the pigeonpea cultivar ICPL 227 and the Rhizobium sp. strain IC 3024 is established, it is efficient in fixing N₂ under saline conditions and can support growth comparable to N-fed plants in growth media with up to 6 dS m^-1 salinity. However, the early stages of establishment of the pigeonpea-IC 3024 symbiotic system have proved sensitive to salinity. The present study showed that the number of nodules was markedly reduced at 8 dS m^-1 salinity; however, nodule development and functioning were not affected by salinity in the pigeonpea-IC 3024 symbiosis. The symbiotic system of Atylosia platycarpa and Rhizobium sp. strain IC 3087 was established successfully even at 12 dS m^-1 and supported growth comparable to that of N-fed plants. P levels in leaves were increased under saline conditions in N-fed and N₂-fixing pigeonpea and A. platycarpa. There were no consistent differences in the leaf Na and chloride levels between N-fed and N₂-fixing plants of pigeonpea and A. platycarpa. The present study suggests that the rhizobial symbiosis may not be a necessary factor for initial screening of pigeonpea and related wild species for salinity tolerance.Submitted as JA No. 964 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Survival and competitiveness of Bradyrhizobium sp. in the rhizosphere of pigeonpea (Cajanus cajan)

Summary The survival of Bradyrhizobium Cajanus strain CC1021 in the soil and rhizosphere of pigeonpea [Cajanus cajan (L.) Millsp] was determined by most probable number and fluorescent antibody techniques. The survival of strain CC1021 was poor under field compared with pot-culture conditions. Although the rhizosphere of pigeonpea promoted the growth of native pigeonpea rhizobia, although no increase in the number of rhizobia was observed with the inoculant strain. Under similar conditions of rhizosphere colonization the competitiveness of strain CC1021 with cv UPAS 120 and ICPL 312 was 10% and 66016 during the first year and 8.4% and 33.3%, respectively, during the second year.     

Variation of morpho-physiological traits in geographically diverse pigeonpea [Cajanus cajan (L.) Millsp] germplasm under different phosphorus conditions

Genotypic variation for morpho-physiological parameters, phosphorus (P) content and root acid phosphatase activity was studied in 52 pigeonpea genotypes. Data related to shoot (length, dry weight, number of leaves, and leaf area), root (volume, length, dry weight, area, perimeter, and number of root tips), acid phosphatase activity, and P content (root, stem, and leaf) were recorded at 60 days after sowing (DAS). The P use efficient genotypes showed high root length, root area, root perimeter, root dry weight, P content in leaves, and root to shoot dry weight ratio under the P not added condition. Significant variation was found among genotypes for root- and shoot-associated characteristics under both P treatments. The P use efficient genotypes with improved root morphological phenes have potential to acquire and utilize more P from immobile soil bound P sources may be of additional factor for increasing efficiency of acquisition and utilization of supplied P fertilizer.     

Establishment and effectiveness of added pigeonpea (Cajanus cajan) rhizobia in different soils of narrow abiotic variability

Summary Pot and laboratory experiments were conducted to study the establishment and effectiveness of a streptomycin-sulphate-resistant (1 mg/ml of medium) pigeonpea rhizobia strain (RM₇) in sterile sand and non-sterile soils. Strain RM₇ increased the drymatter yield of pigeonpea plants (Cajanus cajan) by 106% over control plants under sterile conditions. However, when the rhizobia strain was introduced into 14 different non-sterile soils with a narrow abiotic variability, the comparable beneficial effect was observed only in one soil inoculated with log 6.70 cells/pot. At this inoculum rate, the percentage increase in yield over control plants varied from –1 to 140 in different soils. Rhizobium (RM₇), applied at log 3.70 cells/pot (3 kg soil), showed less than 5% establishment in four soils. However, establishment varied from 8% to 72% at a higher level of inoculation (log 6.70 cells/pot). Displacement of native rhizobia and creation of new sites for nodulation by the introduced rhizobia were also affected by soil properties. The increase in shoot dry-matter yield compared with control plants was positively correlated with the percent establishment of RM₇ (r = 0.60^*) in these soils. Experiments showed that some biotic stresses led to poor survival, proliferation and establishment of the added alien in the soil. Therefore, any culture that is efficient in one soil may not produce similar results under all situations.     

Rhizobium sp. exopolysaccharide production,nodulation, and acetylene reduction activity (N2-fixation) in pigeon pea (Cajanus cajan L.)

Summary There was no correlation between the quantity of exopolysaccharide produced and acetylene reduction activity by Rhizobium spp. or by Bradyrhizobium spp. (Cajanus). The exopolysaccharide-defective mutants of Rhizobium sp. strain P 116 either failed to nodulate or showed a decrease in effectiveness. The deficiency in exopolysaccharide production was corrected by the addition of purified exopolysaccharide from the parent strain, or from Bradyrhizobium sp. strain P 149 or S24 isolated from pigeonpea (Cajanus cajan) and mungbean (Vigna radiata), respectively. However, the nodules so formed were not fully effective compared to those formed by the parent strain.     

Seedling characteristics and retention of current photosynthates in leaves in relation to initial growth in pigeonpea (Cajanus cajan L. Millsp.) and cowpea (Vigna sinensis Endl.)

Improved varieties of pigeonpea (Cajanus cajan L. Millsp.) still retain some growth characteristics of perennial types of wild species such as slow seedling growth and rato on ability which are considered to be major yield-restricting characteristics when the growth period is shortened. Studies on the physiological background of slow initial growth may help improve the potential productivity of short-duration pigeonpea. It was found that the low seedling vigor of pigeonpea could be ascribed to the small seed size (Narayanan et al. 1981; Brakke and Gardner 1987), low rate of photosynthetic CO₂ fixation (Rawson and Constable 1981), and low rates of metabolic processes (Huber and Hanson 1992; Ito et al. 1996). However, few authors have related the slow growth to the retention of current photosynthates and leaf respiration during the night. In contrast, seedling growth of cowpea (Vigna sinensis Endl.) was reported to be vigorous (Brakke and Gardner 1987).

The objectives of this study were to characterize the initial growth of pigeonpea in comparison with cowpea. Two experiments were carried out, one dealing with growth analysis and the other with O2 uptake of leaf and retention of current photosynthates during the night.     

Patterns of diversity in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.) germplasm collected from different elevations in Kenya

Pigeonpea germplasm accessions collected from low (<500 m), medium (501–1000 m), high (1001–1500 m) and very high elevation zones (>1500 m) of Kenya were evaluated for 15 agronomic traits and seed protein content at ICRISAT, Patancheru, India. There were significant differences (P < 0.001) among elevation zones for the number of primary and secondary branches, days to 75% maturity, pod length, seeds per pod, 100-seed weight and seed yield. Mean values indicated that the accessions from low elevation zone were significantly different from those collected in higher elevation zones for early flowering and maturity, number of primary branches, pod length, number of pods per plant, seeds per pod, 100-seed weight, seed yield and harvest index. None of the accessions collected in Kenya belonged to extra early (<80 days to 50% flowering) and early (80–100 days to 50% flowering) maturity groups, as defined by time to flowering at Patancheru, India. Mean diversity index based on all characters indicated that accessions from the low elevation zone are more diverse than those from the higher elevation zones. Frequency distribution for trait extremes indicated that the accessions from the low elevation zone were early to flower and mature, short statured, produced more primary and secondary branches with high pod bearing length, long pods, more pods per plant, more seeds per pod, a high seed yield and harvest index. Accessions from the very high elevation zone were late flowering, with a large number of tertiary branches, large seeds and a high shelling percentage and could be a source for cold tolerance and the breeding of vegetable types. Results suggest that the elevation of collection sites is therefore a very important determinant of variation patterns of pigeonpea in Kenya.     

AFLP analysis in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.) revealed close relationship of cultivated genotypes with some of its wild relatives

A total of 561 amplified fragment length polymorphism (AFLP) loci were generated and used to study the genetic diversity of wild and cultivated genotypes of pigeonpea. Out of 561 marker loci, 558 were polymorphic with an average of 76.12 bands. Analysis of molecular variation (AMOVA) revealed significant strong population structure when genotypes were structured according to continent of origin (F_ST = 0.22) also when structured into cultivated and wild genotypes (F_ST = 0.16). Maximum polymorphic loci were observed in cultivated species C. cajan (352) which is due to more number of genotypes used while, a minimum number of 45 polymorphic bands were obtained in C. acutifolius. Highest (0.291) average gene diversity was recorded in species C. mollis and lowest (0.079) average gene diversity was recorded in C. acutifolius. The 33 genotypes grouped into 14 clusters at 26% Jaccard’s similarity coefficients. Clustering analysis revealed most cultivated genotypes grouped into one major cluster while, the wild genotypes grouped into many clusters at 26% Jaccard’s similarity revealing greater diversity within wild species as compared to cultivated genotypes. From among the various cultivated genotypes studied, four genotypes BRG 3, ICP 7035, TTB 7 and ICP 8863 were molecularly and morphologically diverse and were used as parental genotypes to study nature of inheritance and to identify markers linked to sterility mosaic disease.     

Differences between Cajanus cajan (L.) Millspaugh and C. cajanifolius (Haines) van der Maesen, the progenitor species of pigeonpea

A study was undertaken to know the difference/diversity between pigeonpea and its closely related wild species C. cajanifolius by studying their morphology, crossability, cytology of the hybrid between the two, and molecular studies. Studies revealed that there are at least 5–6 traits that separate the two species such as flower morphology, pod color and morphology, pod constriction, seed color and strophiole, 100 seed weight that separate C. cajan from C. cajanifolius. Molecular studies revealed that a genetic dissimilarity index value ranging from 0.81 to 0.94 exists between the two species.     

Analysis of morphological variation in wild beet (Beta vulgaris L.) from Sicily

Summary In this paper the variation pattern of the Sicilian Beta germplasm collection is analysed. Accessions from Sicily were grown under controlled conditions and were evaluated morphometrically. Geographical distribution patterns of morphological characters are identified and the putative classification of the collection into coastal and inland populations is reassessed. Between adjacent populations significant variation was found for Petiole length, Leaf length, Leaf width and Biomass, however no one variable unambiguously reflected a geographical pattern. Overall differences between coastal and inland populations were small. Petiole length and rate of generative development were found to be important for distinguishing between these groups. Interferences are drawn on how the collection from Sicily could be rationalized to avoid excessive duplication.     

Seed protein diversity in Solanum melongena L. and its wild and weedy relatives

Seed protein profiles by SDS-PAGE were studied in 72 accessions ofSolanum melongena L. and its related taxa in subgenusLeptostemonum (Dunal) Bitter. Comparisons based onJaccard's similarity and UPGMA clustering revealed interrelationships broadly inconformity with the conventional taxonomic treatments. Thus,S. viarum Dunal (sectionAcanthophora Dunal), S.torvum Swartz (section Torva Nees) andS. sisymbriifolium Lam. (sectionCryptocarpum Dunal) were located on separate branches ofthe dendrogram while all taxa of section Melongena Dunal,except S. macrocarpon Dunal, wereclustered closer together. S.melongena, incanum and insanum,all members of the eggplant complex, were revealed to bear very high similaritywith each other. Most of their accessions had identical band patterns supportingthe contention that these represent an interbreeding complex with limitedgenetic differentiation.     

Relationships of Vigna unguiculata (L.) Walp., V. vexillata (L.) A. Rich. and species of section Vigna based on isozyme variation

Summary Isozyme variation in 25 accessions of wild and cultivated Vigna unguiculata, 49 accessions of seven wild species belonging to section Vigna, and 11 accessions of V. vexillata (subgenus Plectrotropis) was scored at 17 putative loci to assess genetic relationships within and among species. The wild species selected for this study are among those which carry important agronomical traits useful in cowpea (V. unguiculata) breeding programs. Allelic frequencies were calculated and Nei's genetic distances were obtained. Low levels of intraspecific variation were observed for V. heterophylla, V. luteola and V. racemosa, whereas the other species showed a higher polymorphism. Vigna unguiculata possessed intraspecific genetic distances comparable to those previously found by other authors. Most of the isozyme variation was apportioned among species. Although V. luteola and V. marina had an interspecific genetic distance resembling the range observed at intraspecific level, all the other species showed very high interspecific distances. Vigna unguiculata was relatively closer genetically to V. vexillata than to the species belonging to section Vigna.Abbreviations AUS Australia - BDI Burundi - BRA Brazil - BWA Botswana - CAF Central African Republic - GHA Ghana - CMR Cameroon - COG Congo - RI Costa Rica - EGY Egypt - ETE Ethiopia - GAB Gabon - GRC Greece - ITA Italy - KEN Kenya - MOZ Mozambique - NER Niger - NGA Nigeria - PAN Panama - RWA Rwanda - TCD Chad - TZA Tanzania - ZAF South Africa - ZAR Zaire - ZMB Zambia     

Characterization and identification of Lactuca sativa cultivars and wild relatives with SDS-electrophoresis (Lactuca sect. Lactuca, Compositae)

Summary Patterns of achene proteins of L. sativa cultivars are mutually compared and compared with L. saligna, L. serriola and L. virosa. L. virosa and L. saligna are easy to identify and are characterized by typical banding patterns. L. sativa and L. serriola share the same banding pattern. They differ clearly from L. saligna and L. virosa. L. sativa and L. serriola are closely related (in terms of similarity), they form a common genepool and L. serriola is involved in the domestication process of L. sativa. Cultivar identification was not possible with the SDS-PAGE method.     

Variation of cultivated flax (Linum usitatissimum L. subsp.usitatissimum) and its wild progenitor pale flax (subsp.angustifolium (Huds.) Thell.)

The domestication of cultivated flax (Linum usitatissimum L. subsp.usitatissimum) is briefly discussed. Using data documented as a matter of routine in genebank work, 63 accessions of cultivated flax from the flax germplasm collection of the Gatersleben Genebank are compared with 73 accessions of its wild progenitor pale flax (subsp.angustifolium (Huds.) Thell.), which have been observed in systematic field trials. Range of variation, genetically based variation, heritability and correlation of several characters are considered, especially with respect to the influence of domestication. Cultivated flax shows higher variation in the characters of generative plant parts, while pale flax varies more in the vegetative parts of the plant. The character correlations are similar in both subspecies. Of the 29 characters studied in pale flax the parameters describing tillering, height of plant, weight of seeds, width of petals and width of leaves are especially suitable for distinguishing between different accessions. In spite of the high heritabilities of most of the characters, the influence of the environment is significant in most cases.