Pod surface exudates of wild relatives of pigeonpea influence the feeding preference of the pod borer, Helicoverpa armigera

Wild relatives of crops are an important source of resistance genes against insect pests. However, it is important to identify the accessions of wild relatives of crops with different mechanisms of resistance to broaden the basis and increase the levels of resistance to insect pests. Therefore, we studied the feeding behavior of pod borer, Helicoverpa armigera, which is the most damaging pest of pigeonpea, in relation to biochemical characteristics of the pod surface exudates in a diverse array of germplasm accessions belonging to 12 species of pigeonpea wild relatives. Feeding by H. armigera larvae was significantly lower on the unwashed or water-, methanol-, or hexane-washed pods of Canajus sericeus, C. scarabaeoides, Flemingia bracteata, F. stricta, and Rhynchosia aurea than those of C. acutifolius, C. albicans, C. cajanifolius, C. lineatus, D. ferruginea, P. scariosa, R. bracteata, and the cultivated pigeonpea, C. cajan genotypes, ICPL 87, and ICPL 332, although there were a few exceptions. The methanol-washed pods of wild relatives were less preferred for feeding by the H. armigera larvae than the unwashed pods, but the hexane-washed pods were preferred more than the unwashed pods. The results suggested that methanol extracted the phagostimulants from the pod surface, while hexane removed the antifeedants. The high-performance liquid chromatography (HPLC) finger printing of methanol and hexane pod surface extracts showed qualitative and quantitative differences in compounds present on the pod surface of different wild relatives of pigeonpea. Some of the peaks in HPLC profiles were associated with feeding preference of the third-instar larvae of H. armigera. There was considerable diversity in wild relatives of pigeonpea as revealed by principal component analysis based on HPLC fingerprints of pod surface extracts in methanol and hexane, and H. armigera feeding on the pods. Wild pigeonpea accessions with low amounts of phagostimulants and high amounts of antifeedants may be used for introgression of resistance genes into the cultivated pigeonpea to develop varieties with broad-based resistance to H. armigera. There is considerable diversity among the wild relatives of pigeonpea, and the accessions with resistance to pod borer. These can be used to broaden the basis and increase the levels of resistance to H. armigera.     

Biosystematic relationships among Cajanus,Atylosia, and Rhynchosia species and evolution of pigeonpea (Cajanus cajan (L.) Millsp.)

Summary Biosystematic studies encompassing morphocytological and electrophoretic analyses of Cajanus cajan, seven species of Atylosia and one of Rhynchosia revealed that A. cajanifolia is closest to C. cajan, followed by A. lineata, A. scarabaeoides, A. sericea, A. albicans, A. volubilis, A. platycarpa and R. rothii, in that order. A revision has been suggested for the taxonomic placement of the seven Atylosia species. Regarding the evolution of cultivated C. cajan, three possible alternatives have been suggested. Firstly, C. cajan could have evolved through gene mutation in A. cajanifolia; secondly, some of the Atylosia species and pigeonpea probably evolved from the same source; and thirdly, the pigeonpea might have developed from naturally occurring interspecific crosses of A. lineata and A. scarabaeoides.     

Morphological and chemical components of resistance to pod borer, Helicoverpa armigera in wild relatives of pigeonpea

Host plant resistance is an important component for minimizing the losses due to the pod borer, Helicoverpa armigera, which is the most devastating pest of pigeonpea. An understanding of different morphological and biochemical components of resistance is essential for developing strategies to breed for resistance to insect pests. Therefore, we studied the morphological and biochemical components associated with expression of resistance to H. armigera in wild relatives of pigeonpea to identify accessions with a diverse combination of characteristics associated with resistance to this pest. Among the wild relatives, oviposition non-preference was an important component of resistance in Cajanus scarabaeoides, while heavy egg-laying was recorded on C. cajanifolius (ICPW 28) and Rhynchosia bracteata (ICPW 214). Accessions belonging to R. aurea, C. scarabaeoides, C. sericeus, C. acutifolius, and Flemingia bracteata showed high levels of resistance to H. armigera, while C. cajanifolius was as susceptible as the susceptible check, ICPL 87. Glandular trichomes (type A) on the calyxes and pods were associated with susceptibility to H. armigera, while the non-glandular trichomes (trichome type C and D) were associated with resistance to this insect. Expression of resistance to H. armigera was also associated with low amounts of sugars and high amounts of tannins and polyphenols. Accessions of wild relatives of pigeonpea with non-glandular trichomes (type C and D) or low densities of glandular trichomes (type A), and high amounts of polyphenols and tannins may be used in wide hybridization to develop pigeonpea cultivars with resistance to H. armigera. Handling editor: Robert Glinwood     

Effect of temperature on seedling growth under impeding conditions

Studies were carried out under controlled laboratory conditions to evaluate the seedling growth capacities of chickpea (Cicer arietinum L.) varieties Pusa 209 and H208 at constant temperatures of 15, 20, 25 and 28°C (±0.5°C) and of pigeonpea (Cajanus cajan L.) variety Prabhat at 20 and 28°C (±0.5°C). Seedling growth at any given time was found to depend on ‘a’, the growth at no impedance, and ‘b’, the impedance growth factori.e., decrease in growth with increase in impedance, and on temperature. The optimum temperature for chickpea was found to be in the range of 20 to 24°C for better seedling growth characteristics, whereas for pigeonpea, 28°C was found to be more congenial than 20°C. Chickpea varieties differed in their response to temperature. Part of the M. Sc. thesis of first author submitted to the Indian Agricultural Research Institute, New Delhi-110012, India.     

Proteinase inhibitors from <Emphasis Type="Italic">Cajanus platycarpus</Emphasis> accessions active against pod borer <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Cajanus platycarpus, a wild relative of Cajanus cajan, is an important source for various agronomically desirable traits, including resistance towards pod borer, Helicoverpa armigera. In the present study, the inhibitory activity of proteinase inhibitors (PIs) present in crude protein extracted from different accessions of C. platycarpus and cultivars of C. cajan was evaluated against H. armigera under in vitro and in vivo conditions. The PIs active against H. armigera gut trypsin-like proteinases (HGPs), referred to as ‘HGPIs’, were more pronounced in mature dry seeds of C. platycarpus accessions when compared with cultivars, which is also evident through gelatin activity staining studies. Therefore, the inhibitory activity of HGPIs was further evaluated in various plant organs of C. platycarpus accessions, such as leaves, flowers, pods, developing seeds at 8–10 days (DAP-I), 18–20 days (DAP-II), and 28–32 days after pollination (DAP-III). However, the HGPI activity was more pronounced in mature dry seeds > DAP-III > DAP-II > DAP-I > flowers > pods > leaves. The observed quantitative allocation of HGPIs closely resembled “Optimal Defense Theory”. Further, bioassays demonstrated that there was a significant reduction in the body weight of the larvae fed upon crude PI extracts of C. platycarpus accessions with concomitant increase in mortality rate and the formation of larval–pupal intermediates. Nevertheless, such changes were not observed when the larvae were fed on crude PI extracts of C. cajan cultivars. These results suggest that the PI gene(s) from C. platycarpus accessions could be exploited in the management of H. armigera by introgression into C. cajan cultivars.     

Trichogramma egg parasitism ofHelicoverpa armigera on short-duration pigeonpea intercultured with sorghum

The levels of egg parasitism byTrichogramma spp. (Hymenoptera: Trichogrammatidae) on populations ofHelicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) were recorded on sorghum (Sorghum bicolor L.) and on two flushes of flowers on short-duration pigeonpea (Cajanus cajan L.).H. armigera oviposition was concentrated on the early flowering stage of sorghum and the flowering and early podding stages of both flushes of pigeonpea. Parasitism on sorghum increased rapidly as egg density increased and reached a peak of 74.6%. Parasitism on pigeonpea was concentrated onH. armigera eggs laid on the first flush of pigeonpea flowers with a maximum of 69.2%. These high levels of parasitism on pigeonpea coincided with the period of parasite activity on sorghum. The levels of parasitism then declined rapidly and only very low levels were detected on a second flush of flowers. This rapid decline resulted in the overall egg mortality caused byTrichogramma on pigeonpea to be low, with a maximum of 7.8% caused by parasitism, compared to 34.4% on sorghum. The pattern of parasitism suggests that a transfer of parasites occurred from sorghum to pigeonpea. The rapid decline of parasitism on the pigeonpea indicates that parasite populations cannot be sustained on pigeonpea once the influx from sorghum stops. The results are discussed in terms of a possible method of encouraging the transfer of parasites from sorghum to short-duration pigeonpea by producing a more continuous cropping environment.     

Egg cannibalism in Helicoverpa armigera on sorghum and pigeonpea

Egg cannibalism by Helicoverpa armigeraHübner (Lepidoptera: Noctuidae) larvae was studied in the laboratory and in the field. In laboratory experiments, first instars were exposed to increasing densities of H. armigera eggs on sorghum, Sorghum bicolor (L.) Moench, and pigeonpea, Cajanus cajan (L.) Millsp. The number of eggs eaten per larva increasedsignificantly as egg availability increased on both sorghum and pigeonpea. In small cages 21–37% of eggs were eaten on sorghum, 4–12%on pigeonpea. Plant feeding declinedsignificantly on both sorghum and pigeonpea asegg density increased. Cannibalism was greateron sorghum than on pigeonpea while plantfeeding was greater on pigeonpea than onsorghum. Only around 8% of eggs were eaten inlarger cages with sorghum. The response toincreasing egg availability in all experimentswas linear. Immunoassay with an anti-vitellinmonoclonal antibody showed that egg cannibalismoccurs on pigeonpea under field conditions.Seven percent of all larvae had egg protein intheir gut. Cannibalism may make a significantcontribution to H. armigera populationsuppression.     

Genome size variation inCajanus cajan (Fabaceae): A reconsideration

A recent investigation of genome size in certain samples of the pigeonpea,Cajanus cajan, indicates values from 1.55 pg to 1.99 pg (1C level), which is 1.29-fold variation between accessions. In the present analysis those of these accessions which had particularly high or low DNA contents in that study were subjected to a reanalysis using propidium iodide and DAPI flow cytometry and Feulgen densitometry. Only minor differences in genome size, not more than 1.047-fold, were found with flow cytometry, and no significant differences were obtained with Feulgen densitometry. The previously reported genome size cannot be confirmed. It is about half as large and was determined in the present study as 0.825 pg (1C, propidium iodide flow cytometry,Glycine max as standard) and 0.853 pg (1C, Feulgen densitometry,Allium cepa andPisum sativum as standards), respectively.     

Genetic variation of trypsin and chymotrypsin inhibitors in pigeonpea [Cajanus cajan (L.) Millsp.] and its wild relatives

Variation in the trypsin inhibitors (TIs) and the chymotrypsin inhibitors (CIs) among 69 pigeonpea [Cajanus cajan (L.) Millsp.] strains from a wide geographical distribution and among 17 accessions representing seven wild Cajanus species was studied by electrophoretic banding pattern comparisons and by spectrophotometric activity assays. The TI and CI electrophoretic migration patterns among the pigeonpea strains were highly uniform but varied in the inhibitor band intensities. The migration patterns of the inhibitors in the wild Cajanus species were highly species specific. The mean TI activity of pigeonpea strains (2279 units) was significantly higher than that of the wild Cajanus species (1407 units). However, the mean CI activity in the pigeonpea strains (62 units) was much lower than that in the wild species (162 units). Kenya 2 and ICP 9151 were the lowest and the highest, respectively, in both the TI and CI activities among all the pigeonpea strains used in this study. A highly-significant positive correlation was observed between the TI and CI activities. The Bowman-Birk type inhibitors with both TI and CI activities were identified in all the pigeonpea strains and also in the accessions of all the wild species except C. volubilis (Blanco) Blanco. The C. volubilis accession ICPW 169 was found to be null for both CI bands and CI activity. Environment, strain, and environment x strain interaction showed highly-significant effects on both the TI and CI activities. Growing the pigeonpea strains at a different environment from their area of adaptation increased TI and CI activities and also altered the maturity period.     

Genetic fingerprinting of pigeonpea [Cajanus cajan (L.) Millsp.] and its wild relatives using RAPD markers

Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of pigeonpea [Cajanus cajan (L.) Millsp.] cultivars and their related wild species. The use of single primers of arbitrary nucleotide sequence resulted in the selective amplification of DNA fragments that were unique to individual accessions. The level of polymorphism among the wild species was extremely high, while little polymorphism was detected within Cajanus cajan accessions. All of the cultivars and wild species under study could be easily distinguished with the help of different primers, thereby indicating the immense potential of RAPD in the genetic fingerprinting of pigeonpea. On the basis of our data the genetic relationship between pigeonpea cultivars and its wild species could be established.NCL Communication No. 6062     

木豆种质资源形态与农艺性状的多样性分析

为挖掘优异木豆(Cajanus cajan)种质资源,对10份木豆种质资源的10个质量性状和18个数量性状的遗传多样性进行了研究,并对其农艺性状进行了聚类分析。结果表明,质量性状的遗传多样性指数均较大,以鲜荚色(1.9219)的最高,其次是旗瓣点缀色、鲜籽粒颜色和干籽粒底色,多样性指数均为1.4855;再次为干籽粒色斑、干籽粒脐环色和有无种阜,均为0.8813;最小的是小叶叶形、旗瓣底色和株型,均为0.7219。聚类分析可将10份木豆种质资源划分为中茎稀疏型、中茎密生型和粗茎密生型3大类型。这为木豆品种选育提供了科学依据。     

A sensitive and rapid immunochemical method for quantitation of proteins

A sensitive and rapid ELISA for quantitation of seed globulins is described. This method employs conjugation of pigeon pea (Cajanus cajan) globulin antibodies and the enzyme peroxidase together with dextran. Using this conjugate, proteins as low as 0.1 ng were detected. Dextran conjugate has a ten-fold greater efficiency of quantitating pigeon pea globulins than the commercial goat anti-rabbit IgG conjugate, and is three-fold more efficient than pigeon pea globulin IgG peroxidase conjugate. The method can be conveniently adapted for quantitation of other proteins also.     

In vitro culture provides additional variation for pigeonpea

Summary The present study is an attempt to exploit somaclonal variation for the varietal improvement of pigeonpea [Cajanus cajan (L) Millsp.]. The pigeonpea plants were regenerated from cotyledon explants, and their progeny was screened for variability. The regenerated R1 plants exhibited a spectrum of alterations in floral morphology and architecture that were absent in the control population. The field-sown R2 plants segregated for traits such as flower color, leaf shape, seed size, color and strophiolation, flowering habit, and fertility. Tissue culture produced different mutational events resulting in both dominant and recessive alleles. Significant variation was observed for plant height, seed mass, and damage due to the insect pest Helicoverpa armigera. The R3 plants, obtained from seed of R2 generation selected for traits such as white seed coat, strophiolation, reduced plant height, seed mass and low damage due to Helicoverpa, maintained the traits when compared with the seed-derived control populations. The results indicate a definite gene for white seed coat and the possibility of additional genes for pest tolerance and high seed mass in an adapted background. Submitted as Journal Article No. 1906 by International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Seedling emergence of chickpea (Cicer arietinum L.), pigeonpea (Cajanus cajan L.) and pearl millet (Pennisetum typhoides L.) Effect of differential soil crusting,as induced by raindrop size,and depth of sowing

The effects of differential soil crusting, as achieved by varying the raindrop size, and depth of sowing on seedling emergence of chickpea (Cicer arietinum L.), pigeonpea (Cajanus cajan L.) and pearl millet (Pennisetum typhoides L.) were investigated. Three drop diameters: 2.75, 1.65 and 0.93 mm and four depths of sowing: 2, 4, 6, and 8 cm were studied. Mean time of emergence increased with increase in raindrop size and depth of sowing. Crops differed in their emergence capacity under adverse soil conditions from delay in the emergence of chickpea to complete failure in the case of pigeonpea. Pearl millet was not significantly affected by drop size but was found to be sensitive to depths beyond 4 cm. Within the chickpea crop, the variety H 208 was observed to have better emergence qualities than Pusa 209. Other parameters like epicotyl diameter, swelling index, and the area of crust broken by the emerging seedling were measured.     

Temperature influence on cohort parameters and demographic characteristics of the two cowpea coreids Clavigralla tomentosicollis and C. shadabi

Age-specific life tables of two important pests of cowpea, Vigna unguiculata (L.) Walp., the pod sucking bugs Clavigralla tomentosicollis Stål and C. shadabi Dolling (Heteroptera: Coreidae), were obtained from observations carried out at different temperatures. A biophysical model was found satisfactory to describe the temperature-response of developmental and mortality rates of egg and nymphal stages, with a peak developmental rate around 34°C in both species. The variability in development times was small and the experimental data did not permit any conclusion with regard to the Erlang probability density function. Survival of eggs and nymphs remained high between 20° and 30°C for both species. At temperatures above 34°C, C. tomentosicollis survivorship and fecundity was higher than that of C. shadabi, which in turn laid more eggs at temperatures between 20° and 30°C. Maximum fecundity is estimated to be at 29°C for C. tomentosicollis (99 eggs/female) and 26°C for C. shadabi (261 eggs/female). At 30°C, the intrinsic rate of increase reached a maximum in both species, 0.152 per day for C. tomentosicollis and 0.145 per day for C. shadabi, and remained high for C. tomentosicollis until 36°C. C. tomentosicollis performed significantly better on pigeonpea, Cajanus cajan Millsp., than on cowpea at higher temperatures.     

Heat Shock Induced Lipid Changes and Solute Leakage in Germinating Seeds of Pigeonpea

Heat shock (HS) reduced total lipid and phospholipid contents and their synthesis in germinating seeds of pigeonpea [Cajanus cajan (L.) Millspaugh]. Lipid peroxidation was also enhanced with increasing temperature and HS duration. HS influenced lipid metabolism to a higher extent at 45°C than at 40°C. This altered lipid metabolism and lipid peroxidation was associated with the loss of various solutes from the germinating seeds, and modification of growth and development. Pretreatment of germinating seeds at 40°C for 1 h or at 45°C for 10 min followed by incubation at 28°C for 3 h prior to 45°C for 2 h ameliorated solute leakage due to reduced lipid peroxidation and improvement in lipid content and membrane function.     

Measurement of N2-fixation in field-grown pigeonpea [Cajanus cajan (L.) Millsp.] using15N-labelled fertilizer

Two experiments were carried out from 1981 to 1983 in Vertisol field at ICRISAT Center, Patancheru, India to measure N₂-fixation of pigeonpea [Cajanus cajan (L.) Millsp.] using the¹⁵N isotope dilution technique. One experiment examined the effect of control of a nodule-eating insect on fixation while another in vestigated the effect of intercroping with cereals on fixation and the residual effect of pigeonpea on a succeeding cereal crop. Although both experiments indicated that at least 88% of the N in pigeonpea was fixed from the atmosphere, one result is considered fortuitous in view of the differential rates of growth of the legume and the control, sorghum [Sorghum bicolor (L.) Moench]. The difference method of calculation in dieated negative fixation and the results emphasized the problem of finding a suitable nonfixing control. In a second experiment, when all plants were confined to a known volume of soil to which¹⁵N fertilizer was added in the field, these problems were overcome, and isotope dilution and difference methods gave similar results of N₂-fixation of about 90%. In intercropped pigeonpea 96% of the total N was derived from the atmosphere. This estimate might be an artifact. There was no evidence of benefit from N fixed by pigeonpea to intercropped sorghum plants. Plant tissue¹⁵N enrichments of cereal crops grown after pigeonpea indicated that the cereal derived some N fixed by the previous pigeonpea. Thus residual benefits to cereals are not only an effect of ‘sparing’ of soil N.     

Cytogenetics of F1 hybrids between Cajanus and Atylosia species and its phylogenetic implications

Summary Cytogenetic relationships between two cultivars of Cajanus cajan and six species of Atylosia were investigated. Of the 12 cross combinations obtained, only seven could be studied. Meiotic chromosome pairing, pollen and ovule fertility in parental species and four F1 hybrids were near normal. Some meiotic abnormalities were observed in the Fls: A. lineata x A. scarabaeoides, A. scarabaeoides x A. sericea and C. cajan (UPAS 120) x A. trinervia, indicating varying degrees of chromosomal and genic differences between these species. These observations suggested that A. cajanifolia is the closest wild relative of C. cajan, followed by A. scarabaeoides, A. albicans and A. trinervia. Among the Atylosia species, A. sericea was closer to A. scarabaeoides than to A. lineata.Botanist, Genetic Resources Unit, ICRISAT, Patancheru-502 324, IndiaRegional Plant Production and Protection Officer, FAO, RAPA, Bangkok-2, Thailand