Characterization of resistance to Callosobruchus maculatus (Coleoptera: Bruchidae) in mungbean variety VC6089A and its resistance-associated protein VrD1

Characteristics of resistance of VC6089A, a mungbean, Vigna radiata (L.) Wilczek, bred by using a wild Vigna species, V. sublobata (Roxburgh) Verdcourt (accession no. TC1966), and containing a novel protein, VrD1, were investigated against the cowpea weevil, Callosobruchus maculatus (F.). The seeds of VC6089A showed high level of resistance; > 96% of the bruchid eggs failed to develop into adults, whereas 85% of eggs laid on susceptible cultivar VC1973A became adults. Mortality of surviving bruchids raised for five generations on VC6089A remained higher than 96%; however, female adults maintained high fecundity and thus showed a positive population growth through these generations. We therefore cannot exclude the possibility that the beetles could develop resistance to the resistant mungbean VC6089A. The protein VrD1 purified from seeds of VC6089A showed marked toxicity to C. maculatus when beetles were reared on artificial seeds containing varying levels of VrD1. Thorough inhibition of development was observed when artificial seeds containing 0.2% (wt:wt) VrD1 was provided for insect feeding. Our findings demonstrated the insecticidal activity of VC6089A mungbean seeds and VrD1 protein against C. maculatus. These results may facilitate safer control against bruchid infestation.     

Vicilin variants and the resistance of cowpea (Vigna unguiculata) seeds to the cowpea weevil (Callosobruchus maculatus)

1. A globulin fraction prepared from the meal of Callosobruchus maculatus-resistant cowpea (Vigna wiguiculata) seeds was shown to be detrimental to this bruchid when incorporated in artificial seeds.2. The performance of C. maculatus was also shown to be strongly hindered by vicilins from resistant seeds when these storage proteins were incorporated in artificial seeds at the level of 2%.3. The purified vicilins from seeds of both resistant and susceptible cowpea varieties were shown to have the same SDS-PAGE pattern but different mobilities in non-denaturing polyacrylamide gel electrophoresis.4. These results and previous ones obtained by us (Silva and Xavier-Filho, 1991; Sales et al., 1992) strongly suggest that the resistance of cowpea seeds from the cultivar TVu 2027 and from others bred from it is associated with the presence of vicilin molecules which are refractory to digestion by bruchid midgut proteinases.     

栽培绿豆V2709抗豆象特性遗传分析

利用原产于印度的抗豆象栽培绿豆品种V2709与农艺性状优良的感豆象推广品种中绿1号(VC1973A)杂交,以杂交种F1、F2为试验材料,通过田间农艺性状调查、室内人工接虫鉴定及多元相关分析等,研究V2709中所包含的抗豆象特性的遗传背景。结果表明V2709的抗豆象特性由一对显性基因控制;通过相关性分析发现其抗虫性与植株高度呈极显著正相关,与主茎节数呈显著正相关,而与其他性状无明显相关性。     

The Effect of Dung and Dispersal on Postdispersal Seed Predation of Attalea phalerata (Arecaceae) by Bruchid Beetles1

Low postdispersal mortality of palm seeds in tapir dung is hypothesized to result from the mechanical barrier provided by dung against bruchid infestation and/or from the distance to adult palms at which seeds are dispersed. We tested these hypotheses by distributing endocarps of Attalea phalerata Mart. ex Spreng. in experimental dung piles in Beni, Bolivia. Predation rates were significantly lower for seeds covered by dung than for exposed or partially covered seeds, but did not differ between seeds placed below and 50 m away from palms. Thus, dung, not short‐distance dispersal, protects seeds against bruchid beetles, and may ultimately promote survival of palm seeds.     

Large African herbivores,bruchid beetles and their interactions with Acacia seeds

This study investigated the interactions of large African herbivores and bruchid seed beetles with Acacia seeds. The germination of bruchid-infested and uninfested seeds was compaed. The effects of pod consumption by large herbivores on bruchid infestation and seed germination was also assessed. Bruchid-infested seeds did germinate, and the germination of bruchid-infested and uninfested A. tortilis, A. nilotica and A. hebeclade seeds did not differ. Pod ingestion by large herbivores lowered the bruchid infestation of consumed and defaecated seeds compared to uningested seeds. Uninfested, ingested and voided A. tortilis seeds germinated seeds. Furthermore, infested A. tortilis seeds egested by giraffe, kudu and ostrich germinated better than infested, uningested seeds. Pod ingestion by large herbivores may reduce bruchid infestation, increase Acacia seed germination and therefore increase potential Acacia seedling recruitment.     

Characterization of resistance to three bruchid species (Callosobruchus spp., Coleoptera,Bruchidae) in cultivated rice bean (Vigna umbellata)

Resistance of wild and cultivated rice bean (Vigna umbellata [Thunberg] Ohwi and Ohashi) to three bruchid species, Callosobruchus chinensis L., Callosobruchus maculatus F., and Callosobruchus analis F., was evaluated. All but three accessions of cultivated, and all wild rice bean accessions tested, exhibited complete resistance to all three bruchid species. Rice bean seeds with seed coat removed also showed complete resistance to the three bruchid species. Results indicate that physical attributes and/or chemical(s) present in the seed coat of rice bean are not the main factors responsible for resistance. Feeding tests were performed by using artificial beans prepared with varying proportions of rice bean (resistant) and azuki bean (susceptible) flour. Number of bruchid adults that emerged decreased, and larval developmental period (days) was extended, when artificial beans with an increasing proportion of rice bean flour were used. These tests revealed that a chemical compound(s) contained in the cotyledon of rice bean has an inhibitory effect on the growth of these bruchid species. The results also indicate that the chemical(s) in rice bean cotyledon is most effective against C. maculatus.     

Defensive Responses in Groundnut Against Chewing and Sap-Sucking Insects

Induced resistance is one of the important components of host plant resistance to insects. We studied the induced defensive responses in groundnut genotypes with different levels of resistance to the leaf defoliator Helicoverpa armigera and the sap-sucking insect Aphis craccivora to gain an understanding of the induced resistance to insects and its implications for pest management. The activity of the defensive enzymes (peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, superoxide dismutase, ascorbate peroxidase, and catalase) and the amounts of total phenols, hydrogen peroxide, malondialdehyde, and proteins were recorded at 6 days after infestation. Induction of enzyme activities and the amounts of secondary metabolites were greater in the insect-resistant genotypes ICGV 86699, ICGV 86031, ICG 2271, and ICG 1697 infested with H. armigera and A. craccivora than in the susceptible check JL 24. The resistant genotypes suffered lower insect damage and resulted in lower Helicoverpa larval survival and weights than those larvae fed on the susceptible check JL 24. The number of aphids was significantly lower on insect-resistant genotypes than on the susceptible check JL 24. The results suggested that groundnut plants respond to infestation by H. armigera and A. craccivora in a similar way; however, the degree of the response differed across the genotypes and insects, and this defense response is attributed to various defensive enzymes and secondary metabolites.     

Resistance of stored bean varieties to Acanthoscelides obtectus (Coleoptera: Bruchidae)

During bean seed storage, yield can be lost due to infestations of Acanthoscelides obtectus Say, the bean weevil. The use of resistant varieties has shown promising results in fighting these insects, reducing infestation levels and eliminating chemical residues from the beans. The expression of resistance to A. obtectus in bean varieties is frequently attributed to the presence of phytohemagglutinins, protease inhibitors and alpha-amylase, and especially to variants of the protein arcelin, which reduce the larval viability of these insects. To evaluate the effect of bean seed storage time on the resistance expression of bean varieties to A. obtectus , tests with seeds of three ages (freshly-harvested, 4-month-old, and 8-month-old) were conducted in the laboratory, using four commercial varieties: Carioca Pitoco, Ipa 6, Porrillo 70, ônix; four improved varieties containing arcelin protein: Arc.1, Arc.2, Arc. 3, Arc.4; and three wild varieties also containing arcelin protein: Arc.1S, Arc.3S, and Arc. 5S. The Arc.5S, Arc.1S, and Arc.2 varieties expressed high antibiosis levels against the weevil; Arc.1 and Arcs expressed the same mechanism, but at lower levels. The occurrence of oviposition non-preference was also observed in Arc.5S and Arc.1S. The Arc.3 and Arc. 4 varieties expressed low feeding non-preference levels against A. obtectus. The expression of resistance in arcelin-bearing, wild or improved varieties was affected during the storage of seeds, and was high under some parameters but low in others. The results showed that addition of chemical resistance factors such as protein arcelin via genetic breeding may be beneficial in improving the performance of bean crops.     

Mapping of quantitative trait loci for a new source of resistance to bruchids in the wild species Vigna nepalensis Tateishi & Maxted (Vigna subgenus Ceratotropis)

Azuki bean breeders have long been interested in producing azuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] varieties with bruchid resistance. A new bruchid (Callosobruchus spp.) resistance source was found in V. nepalensis Tateishi & Maxted, a species that is cross compatible with azuki bean. Quantitative trait loci (QTLs) analysis for resistance to C. chinensis (L.) and C. maculatus (F.) was conducted using F(2) (V. nepalensis x V. angularis) and BC(1)F(1) [(V. nepalensis x V. angularis) x V. angularis] populations derived from crosses between the bruchid resistant species V. nepalensis and bruchid susceptible species V. angularis. Resistance was measured using two traits, percentage of seeds damaged by bruchids and the time taken for adult bruchids to emerge from seeds. Based on the results from both populations seven QTLs were detected for bruchid resistance; five QTLs for resistance to C. chinensis and two QTLs for resistance to C. maculatus. The different locations found for some resistance QTL to the two bruchid species suggests different resistance mechanisms. QTLs on linkage group (LG) 1 and LG2 for bruchid resistance to C. chinensis co-localized with seed size QTLs suggesting that incremental increase in seed size accompanied susceptibility to C. chinensis. Based on linked markers the QTL on these two linkage groups appear to be the same as previously reported in other Asian Vigna. However, several other QTLs were newly detected including one on LG4 that appears unrelated to seed size. Transfer of these new sources of bruchid resistance from V. nepalensis to azuki bean will be aided by the progress being made in azuki genome mapping.     

马尾松受害诱导的化学物质滞后变化

通过接种马尾松毛虫Dendrolimus punctatus和人工剪叶,分析比较了不同受害方式对马尾松针叶内化学物质的影响。结果表明,害虫取食松针诱导的次生代谢物质(单宁、酚类物质)比人工剪叶处理略有增加,而营养物质-可溶性糖变化不大。对马尾松进行害虫取食为害、人工剪叶受害、未受害三种处理后,连续3年跟踪测定了松针叶内化学物质含量的变化。结果发现,无论是害虫取食为害,还是人工剪叶受害,针叶内次生代谢物质含量都减少,可溶性糖含量降低,直至一年后才恢复到原有的水平;蛋白质变化的趋势则始终是受害处理的含量比未受害处理的含量高,表明马尾松受害后诱导的化学物质变化具有滞后特性。     

Effects of seed type and bruchid genotype on the performance and oviposition behavior of Zabrotes subfasciatus (Coleoptera: Bruchidae)

The effect of different bean varieties on the performance of the bruchid beetle Zabrotes subfasciatus Boheman (Coleoptera: Bruchidae), was determined by using wild and cultivated seeds of the genus Phaseolus. Results showed that the quality of the host plant affected the performance and the oviposition behavior of female beetles. Overall, bruchid performance was higher on cultivated seeds than on wild seeds. It was also found that the oviposition behavior and the performance of their offspring differed between females that originated from wild versus cultivated seeds. We also demonstrated the importance of a genetic component in bruchid performance: longevity, fecundity, larval development, adult size, and the sex ratio of the progeny varied according to their maternal lineage (‘familyrsquo;). For example, on the same host type, some females laid twice as many eggs as females from other ‘familiesrsquo;, and the sex ratio among the offspring correlated with that of their mother. Thus, the performance and behavior of Z. subfasciatus are not only affected by environmen tal factors such as the quality of the seeds on which they develop, but also have a genetic basis which can counterbalance a less suitable quality of the host plant. For a crop pest such as Z. subfasciatus, its ability to survive and adapt on host plants of differing quality may be an important attribute to consider for pest management.     

Three–way interactions between Acacia, large mammalian herbivores and bruchid beetles - a review

Large mammalian herbivores are both predators and dispersers of Acacia seeds. While some of the seeds are destroyed during passage through the herbivore's digestive tract, others are defecated unharmed. Ingestion by large herbivores facilitates germination by scarification of the seed coat. The extent of the influence of herbivores on seed dispersal and germination depends on seed retention time and tooth size, which are both positively correlated with body size. Infestation by bruchid beetles (Bruchidae) reduces Acacia germination. Herbivores may reduce bruchid infestation in several ways. Larvae in recently infested seeds are killed by stomach acids penetrating the seed through the larval entry hole. Seeds that are partly excavated by burrowing larvae in more advanced stages may be crushed by the herbivore's teeth. Lastly, but probably most crucially, herbivores simply remove seeds from the natal tree prior to infestation or at least prior to reinfestation. The timing and magnitude of herbivory is crucial for both the reduction of bruchid infestation and Acacia seedling establishment. Although it is widely agreed that a three–way interaction exists between bruchid beetles, Acacia trees and large mammalian herbivores, it is also apparent that the relationship is highly complex and is not yet completely understood.     

Revealing different systems responses to brown planthopper infestation for pest susceptible and resistant rice plants with the combined metabonomic and gene-expression analysis

Brown planthopper (BPH) is a notorious pest of rice plants attacking leaf sheaths and seriously affecting global rice production. However, how rice plants respond against BPH remains to be fully understood. To understand systems metabolic responses of rice plants to BPH infestation, we analyzed BPH-induced metabolic changes in leaf sheaths of both BPH-susceptible and resistant rice varieties using NMR-based metabonomics and measured expression changes of 10 relevant genes using quantitative real-time PCR. Our results showed that rice metabonome was dominated by more than 30 metabolites including sugars, organic acids, amino acids, and choline metabolites. BPH infestation caused profound metabolic changes for both BPH-susceptible and resistant rice plants involving transamination, GABA shunt, TCA cycle, gluconeogenesis/glycolysis, pentose phosphate pathway, and secondary metabolisms. BPH infestation caused more drastic overall metabolic changes for BPH-susceptible variety and more marked up-regulations for key genes regulating GABA shunt and biosynthesis of secondary metabolites for BPH-resistant variety. Such observations indicated that activation of GABA shunt and shikimate-mediated secondary metabolisms was vital for rice plants to resist BPH infestation. These findings filled the gap of our understandings in the mechanistic aspects of BPH resistance for rice plants and demonstrated the combined metabonomic and qRT-PCR analysis as an effective approach for understanding plant-herbivore interactions.     

Survival and Scatterhoarding of Frugivores-Dispersed Seeds as a Function of Forest Disturbance

The effect of forest disturbance on survival and secondary dispersal of an artificial seed shadow (N= 800) was studied at Brownsberg Natural Park, Suriname, South America. We scattered single seeds of the frugivore‐dispersed tree Virola kwatae (Myristicaceae), simulating loose dispersal by frugivores, in undisturbed and disturbed secondary forest habitats. Seed survival rate aboveground was high (69%) within 2 wk and was negatively correlated with scatterhoarding rate by rodents, the latter being significantly lower in the undisturbed forest (9%) than in the disturbed forest (20%). Postdispersal seed predation by vertebrates was low (3%) and infestation of seeds by invertebrates was almost zero in all instances. Therefore, secondary seed dispersal by rodents in forest is not as critical for recruitment as observed among other bruchid‐infested large‐seeded species. Secondary seed dispersal by rodents may, however, facilitate seedling recruitment whether cached seeds experience greater survival than seeds remaining above ground surface.     

Molecular markers linked with bruchid resistance in Vigna radiata var. Sublobata and their validation

Bruchid, Callosobruchus chinensis (L.) is an important pest of Vigna radiata during storage. RFLP and PCR based markers identified, linked with bruchid resistance gene in wild accession of greengram (V. radiata var. Sublobata) either collected from Madagaskar or Australia. Whether these markers will be useful for marker assisted introgression of bruchid resistance gene from the Indian accession into the existing cultivars are not known. Here, we employed two STS based markers which were found earlier, to be linked with bruchid resistance gene in Australian accession ACC41. Only one primer pair, STSbr1 showed polymorphism among Indian Sublobata accession (Sub2) and other twelve green gram cultivars. Analysis of 113 segregating lines (F₆) of a cross between a popular cultivar of West Bengal, B1 and Sub2 showed a cent percent co-segregation of resistant locus with the polymorphic fragment. STSbr1 behave as a dominant marker among Indian genotypes although it has been shown earlier a co-dominant banding pattern between ACC41 and other Australian Susceptible cultivars. Other STS marker, STSbr2, does not produce any polymorphic fragment among Sub2 and 18 greengram genotypes. STSbr1 employed in screening of 50 green gram accessions and found high efficiency in screening of bruchid resistant genotypes also. So STSbr1 will be useful for marker assisted selection and germplasm screening for development of bruchid resistant greengram.     

Maternal effects on the life histories of bruchid beetles infesting Acacia raddiana in the Negev desert, Israel

Acacia trees (Leguminosae) in the Negev desert of Israel are in substantial danger of extinction as a result of high mortality, caused by anthropogenically induced water stress and very low recruitment, which is highly negatively affected by bruchid beetle infestation. Intensive seed infestation (up to 97%) by bruchids on Acacia species in the Negev desert has been described. We hypothesized that water‐stressed trees would be less able to produce secondary defense compounds in their seeds to decrease seed herbivory and hence suffer higher infestation. Thus, there should be a negative correlation between the beetle's fitness and the tree's physiological state. We further hypothesized the mechanism of the correlation to be maternal effects on beetle quality. Thus, beetles whose mothers were reared on seeds of trees in a poor state should have higher fitness, regardless of the quality of the seeds in which they were reared. We reared F₁ generation Caryedon palaestinicus Southgate (Coleoptera: Bruchidae) in seeds of Acacia raddiana Savi (Leguminosae) trees in good and in poor physiological state. We then conducted a reciprocal infestation experiment enabling females to lay on seeds from both sources. We found maternal effects on offspring number but not on body mass. The major effect was the limited survival of maternal beetles on trees in good physiological state. However, contrary to our prediction, C. palaestinicus developing in seeds from Acacia raddiana in good physiological state had significantly higher body mass.     

Variation in neurotoxin, trypsin inhibitors and susceptibility to insect attack in varieties of Lathyrus sativus seeds.

Ten varieties of Lathyrus sativus seeds were analysed for the neurotoxin (BOAA) and trypsin inhibitor (TI) content. The BOAA content varied from 0.142 to 0.680%. The values of trypsin inhibitor content were also uniform with the exception of two varieties whose values were comparatively low. When the seeds were exposed to insects all the varieties were affected, but the two varieties showing low trypsin inhibitor content also showed some protection against infestation. Variety P-24 was low in BOAA and trypsin inhibitor content with comparatively low insect susceptability.     

Aphid infestation promotes survival of a seed predator: observations and experiments on a tritrophic community module

We investigated whether aphid presence and abundance influence the survival of an endophagous pre-dispersal seed predator of the same host plant. We studied a terrestrial community module consisting of one plant (Laburnum anagyroides) and four insect species/groups (an aphid, Aphis cytisorum, a pre-dispersal seed predator bruchid, Bruchidius villosus, aphid-attending ant species, and parasitoids of the bruchid). Two complementary investigations were carried out in parallel: (a) a plant-aphid-ant complex was experimentally manipulated by excluding aphids, ants, or both for 5 years to assess their impacts on the seed predator’s survival and parasitism rate; and (b) different aphid infestation levels on randomly selected infructescences were correlated with plant traits, nutrient allocation pattern, and variables of seed predator’s survival, such as the number of eggs laid and adults emerged influenced by parasitoid activity, for 7 years. We found that ants did not affect bruchid oviposition negatively, but egg-parasitism was significantly decreased by their presence. Plant traits, such as the number of seeds and seed mass, as well as seed predator performance were negatively affected by heavy aphid infestation. Seed predator -infested seeds had no effect on the mass of remaining seeds in the pods. This study suggests that aphids were nevertheless promoting bruchid abundance and survival, depending on their infestation rate.     

Relationship Among Phenolic Contents,Seed Predation,and Physical Seed Traits in <Emphasis Type="Italic">Mimosa bimucronata</Emphasis> Plants

Phenolic contents were compared between Mimosa bimucronata seeds from infested and non-infested fruits to assess induced defense response. By measuring leg length of the bruchid beetle Acanthoscelides schrankiae, we verified whether phenolic contents affected bruchid body size. In addition, the relationship between physical seed traits and phenolic contents was examined. Results showed that seeds from infested fruits had significantly greater phenolic contents than seeds from non-infested fruits, which suggested induced defense. Body size variation in A. schrankiae was marginally nonsignificant according to phenolic contents among plants (negative trend), indicating that phenols may interfere directly with bruchid performance. Seeds that were more irregularly shaped had significantly greater phenolic contents than those that were more uniform. Therefore, the most perfectly spherical seeds may be more vulnerable to seed predation, and our results suggest that the production of phenolic compounds was increased in infested fruits, which in turn may affect A. schrankiae development.     

An eco-metabolomic study of host plant resistance to Western flower thrips in cultivated,biofortified and wild carrots

Domestication of plants and selection for agronomic traits may reduce plant secondary defence metabolites relative to their ancestors. Carrot (Daucus carota L.) is an economically important vegetable. Recently, carrot was developed as a functional food with additional health-promoting functions. Biofortified carrots contain increased concentrations of chlorogenic acid as an antioxidant. Chlorogenic acid is involved in host plant resistance to Western Flower Thrips (Frankliniella occidentalis), one of the key agri- and horticultural pests worldwide. The objective of this study was to investigate quantitative host plant resistance to thrips in carrot and to identify candidate compounds for constitutive resistance. As such we explored whether cultivated carrot is more vulnerable to herbivore attack compared to wild carrot. We subjected a set of 14 biofortified, cultivated and wild carrot genotypes to thrips infestation. We compared morphological traits and leaf metabolic profiles of the three most resistant and susceptible carrots using nuclear magnetic resonance spectroscopy (NMR). In contrast to our expectation, wild carrots were not more resistant to thrips than cultivated ones. The most thrips resistant carrot was the cultivar Ingot which is known to be tolerant against carrot root fly (Psila rosae). Biofortified carrots were not resistant to thrips. Plant size, leaf area and number of leaf hairs did not differ between resistant and susceptible carrots. The metabolic profiles of the leaves of resistant carrots were significantly different from those of susceptible carrots. The leaves of resistant carrots contained higher amounts of the flavanoid luteolin, the phenylpropanoid sinapic acid and the amino acid β-alanine. The negative effect of these compounds on thrips was confirmed using in-vitro bioassays. Our results have potential implications for carrot breeders. The natural variation of metabolites present in cultivated carrots can be used for improvement of thrips resistance. This is especially promising in view of the candidate compounds we identified since they do not only confer a negative effect on thrips but as antioxidants also play an important role in the improvement of human health.