Free amino acid content of phloem sap from Brassica plants in relation to performance of Lipaphis erysimi (Hemiptera: Aphididae)

Cultivars of Brassica juncea, B. napus and B. campestris, differing in host-suitability to the mustard aphid (Lipaphis erysimi pseudobrassicae Kalt.), were analysed for their phloem sap content of free amino acids. Sap was collected from excised mustard aphid stylets and analysed by means of high performance liquid chromatography. Aspartic and glutamic acid were, together with glutamine, the dominating amino acids. All plants had very little glycine, γ-aminobutyric acid (GABA), methionine and tryptophan in their sap. This amino acid pattern closely resembled that of cereals. The total concentration and the individual composition of amino acids were not found to be related to the level of aphid resistance previously found in the investigated Brassica species. However, one B. campestris cultivar, yellow sarson YSB-9, on which the aphids were shown to grow more slowly, had less amino acid content than other B. campestris varieties. The slow population growth of aphids on B. juncea, compared to that on B. campestris, is probably due to other than nutritional factors.     

Assessment of aphid infestation levels in some cultivars of mustard with varying defensive traits

Phloem-sucking mustard aphid (Lipaphis erysimi) is a major pest of mustard (Brassica juncea). Pot experiment with randomised block design was conducted with five replicates of each of five cultivars (Alankar, Pusa Jai Kisan, Rohini, Sakha and Varuna) of the mustard for their degree of inherited resistance and/or susceptibility to the mustard aphid infestation. Forty-five days old (from date of sowing) pot-grown plants of all selected cultivars of mustard were exposed to 40 adult mustard aphids. The aphid-infested plants were kept in specially designed net houses of fine mesh to protect from predators and/or migration of aphids from one to other host. The aphid population and some growth attributes of the selected cultivars of mustard were recorded 15 and 30 days later (i.e. at 60 and 75 days after sowing). The aphid population multiplied more rapidly on Rohini than other four cultivars. Cultivar Alankar resisted most and supported to least number of aphid’s off-springs. Statistically analysed growth attributes (fresh plant mass, dry plant mass, protein, chlorophyll and carotenoid contents), resistance attribute (proline) and population demography of aphids revealed that some inherited characteristics of avoidance, antibiosis and repellence to herbivores helped cultivar Alankar to excel despite equal degree of aphid attacks as on other cultivars. Cultivar Rohini for the want of such resisting factors remained vulnerable to aphid herbivory. These two cultivars (Alankar and Rohini) form good research material for comparative studies on plant defences to herbivory and a tri- trophic resistance through volatile chemical signalling.     

Elicitation of jasmonate-mediated host defense in Brassica juncea (L.) attenuates population growth of mustard aphid Lipaphis erysimi (Kalt.)

The productivity of Brassica oilseeds is severely affected by its major pest: aphids. Unavailability of resistance source within the crossable germplasms has stalled the breeding efforts to derive aphid resistant cultivars. In this study, jasmonate-mediated host defense in Indian mustard Brassica juncea (L.) Czern. was evaluated and compared with regard to its elicitation in response to mustard aphid Lipaphis erysimi (Kalt.) and the defense elicitor methyl jasmonate (MeJ). Identification of jasmonate-induced unigenes in B. juncea revealed that most are orthologous to aphid-responsive genes, identified in taxonomically diverse plant–aphid interactions. The unigenes largely represented genes related to signal transduction, response to biotic and abiotic stimuli and homeostasis of reactive oxygen species (ROS), in addition to genes related to cellular and metabolic processes involved in cell organization, biogenesis, and development. Gene expression studies revealed induction of the key jasmonate biosynthetic genes (LOX, AOC, 12-OPDR), redox genes (CAT3 and GST6), and other downstream defense genes (PAL, ELI3, MYR, and TPI) by several folds, both in response to MeJ and plant-wounding. However, interestingly aphid infestation even after 24 h did not elicit any activation of these genes. In contrast, when the jasmonate-mediated host defense was elicited by exogenous application of MeJ the treated B. juncea plants showed a strong antibiosis effect on the infesting aphids and reduced the growth of aphid populations. The level of redox enzymes CAT, APX, and SOD, involved in ROS homeostasis in defense signaling, and several defense enzymes viz. POD, PPO, and PAL, remained high in treated plants. We conclude that in B. juncea, the jasmonate activated endogenous-defense, which is not effectively activated in response to mustard aphids, has the potential to reduce population growth of mustard aphids.     

植物提取物对萝卜蚜的室内毒力研究

采用浸叶法和Potter喷雾法测定七种植物提取物对萝卜蚜 (Lipaphiserysim)的室内毒力效果。研究结果显示 :巴豆、博落回和醉鱼草提取物对萝卜蚜的致死中浓度LC50 分别为 1 86.94mg/L、2 0 0 .75mg/L和 1 5 7.73mg/L。     

Host quality of different aphid species for rearing Diaeretiella rapae (McIntosh) (Hymenoptera: Braconidae)

This study aimed to evaluate the quality of the aphid Myzus persicae (Sulzer), Lipaphis erysimi (Kaltenbach) and Brevicoryne brassicae (L.) as hosts for the parasitoid Diaeretiella rapae (McIntosh). Parasitization by D. rapae was higher on M. persicae than on L. erysimi and B. brassicae. The time of development of D. rapae from egg to mummy or egg to adult male or female were shorter on M. persicae than on L. erysimi and B. brassicae. Moreover, D. rapae showed no significant differences in the emergence rate, sex ratio and longevity when reared on the three aphid species. Myzus persicae was the largest aphid host, with B. brassicae and L. erysimi being of intermediate and of small size, respectively. Diaeretiella rapae reared on M. persicae was larger than when reared on L. erysimi and B. brassicae, and females of D. rapae were significantly larger than males on M. persicae, but males of D. rapae were larger than females when reared on L. erysimi. No difference in size was detected between males and females in parasitoids reared on B. brassicae. Among the aphid species studied, M. persicae was found to be the most suitable to D. rapae.     

桃蚜、萝卜蚜的种内密度和种间竞争效应

在温室内不同密度条件下,研究了桃蚜和萝卜蚜在甘蓝上的种内密度效应和种间竞争作用。结果表明：单独饲养时,在5头／株和10头／株密度下,两种蚜虫种内密度效应均较弱;而在15头／株时,桃蚜和萝卜蚜的寿命和产蚜量都随之显降低,说明当两种蚜虫达到一定的拥挤程度时,种内密度对种群均产生一定的负效应。两种蚜虫共存时,单头产蚜量均比单独饲养时显下降,种问竞争作用明显。在10、15头／株密度下,桃蚜的竞争作用大于萝卜蚜,萝卜蚜的寿命和单头产蚜量都极显低于桃蚜。     

多种植物提取物对桃蚜和萝卜蚜的拒食活性研究

采用改进的蜜露色斑法 ,测试 34种植物乙醇提取物 (0 0 1g干重·mL-1,0 0 1gDW·mL-1)和几种天然源药剂对桃蚜Myzuspersicae(Sulzer)和萝卜蚜Lipaphiserysimi(Kaltenbach)取食的影响。结果表明 ,白蝴蝶Syngonium podophyllumSchott.、苍耳XanthiumsibiricumPrtr.etWidd、白花非洲山毛豆Tephrosiavo gelliHook .的提取物对两种蚜虫的活性均较高 ,2 4h对桃蚜的拒食率分别达 0 6 82、0 6 2 9和 0 80 6 ,对萝卜蚜的拒食率分别为 0 4 0 5、0 370和 0 383,与对照之间差异极显著 ;油茶Camelliaoleifera和花椒Zanthoxylumschinifolium的提取物拒食作用亦较强 ,2 4h对桃蚜的拒食率分别为 0 5 5 2和 0 5 14 ,对萝卜蚜的拒食率分别为 0 5 70和 0 5 18。三种药剂 (0 3%印楝素乳油、鱼藤酮精、机油乳剂 )在所测试的浓度对两种蚜虫的拒食率均达 0 719以上 ,拒食效果显著。     

Positive correlation of methamidophos resistance between Lipaphis erysimi and Diaeretilla rapae and effects of methamidophos ingested by host insect on the parasitoid

Temporal and spatial correlated variations on the methamidophos resistance and biomolecular rate constant of acetylcholinesterase to insecticides were found between the turnip aphid, Lipaphis erysimi and its endoparasitoid, Diaeretilla rapae , collected from field colonies and an insecticide-free field insectarium in Fujian, China. Compared to the related susceptible insectarium population, L. erysimi and D. rapae displayed 7.4–29.2- and 2.6–9.2-fold resistance ratios, respectively. In addition, two populations of L. erysimi with different methamidophos resistance levels, that is, a field (with 5.8-fold resistance ratio) and an insectarium population, were used to study the effects of methamidophos ingested by the host insect on D. rapae development. The percentage of D. rapae cocoon formation decreased significantly when the parasitized L. erysimi were fed on cauliflower leaves treated with methamidophos at lethal concentration dosages 10 (LC₁₀) or LC₅₀. At LC₅₀ dosages the percentage of D. rapae cocoon formation and adult emergence decreased significantly. When the parasitized L. erysimi were fed on methamidophos at LC₉₀ dosage, no D. rapae cocoons were found. When the field or insectarium L. erysimi were treated with methamidophos at LC₁₀, the susceptibility to methamidophos in the adult D. rapae emerged from the treated host insect was similar to the control. However, the susceptibility to methamidophos in the adult D. rapae became lower than the control when the host insects were treated at LC₅₀ dosages. The data thus suggested that the methamidophos ingested by the host insect L. erysimi could be an important factor in the endoparasitoids' insecticide resistance development. The natural selectivity would favor the parasitoids that had developed an insensitivity to the insecticide(s).     

High efficiency in planta transformation of Indian mustard (Brassica juncea) based on spraying of floral buds

Plant Cell, Tissue and Organ Culture (PCTOC) - Brassica juncea (Indian mustard) and its sub-varieties include whole range of oilseed and vegetable mustard in China, Canada, Australia, Europe and...     

Mutant acetolactate synthase gene is an efficient in vitro selectable marker for the genetic transformation of Brassica juncea (oilseed mustard)

We report in this study, the successful deployment of a double mutant acetolactate synthase gene (ALSdm, containing Pro 197 to Ser and Ser 653 to Asn substitutions) as an efficient in vitro selection marker for the development of transgenic plants in Brassica juncea (oilseed mustard). The ALS enzyme is inhibited by two categories of herbicides, sulfonylureas (e.g. chlorsulfuron) and imidazolinones (e.g. imazethapyr), while the mutant forms are resistant to the same. Three different selection agents (kanamycin, chlorsulfuron and imazethapyr) were tested for in vitro selection efficiency in two B. juncea cultivars, RLM198 and Varuna. For both the cultivars, higher transformation frequencies were obtained using chlorsulfuron (3.8 +/- 0.6% and 4.6 +/- 0.9% for RLM198 and Varuna, respectively) and imazethapyr (10.2 +/- 0.7% for RLM198 and 7.8 +/- 1.2% for Varuna) as compared to that obtained on kanamycin (3.1 +/- 0.2% and 2.8 +/- 0.5% for RLM198 and Varuna, respectively). Additionally, transformation frequencies were higher on imazethapyr than on chlorsulfuron for both the cultivars indicating that imidazolinones are better selective agents than sulfonylureas for the selection of mustard transgenics.     

Immature stages of some aphidophagous syrphid flies of India (Insecta, Diptera, Syrphidae)

Immature stages, viz. egg, larva and puparium, of six species of the economically most important syrphid flies, Scaeva latimaculata (Brunetti), Ischiodon scutellaris (Fabricius), Episyrphus altemans (Macquart), Sphaerophoria Indiana Bigot, Metasyrphus confrater (Wiedemann), and M. latilunulatus (Collin), have been studied. These species are voracious feeders on three aphid species, Lipaphis erysimi (Kaltenbach), Myzus persicae (Sulzer) and Brevicoryne brassicae (L.), which are serious pests of mustard, Brassica campestris L.     

Transgenic Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) expressing tomato glucanase leads to arrested growth of <Emphasis Type="Italic">Alternaria brassicae</Emphasis>

Brassica juncea is an important oilseed crop of the Indian sub-continent. Yield loss due to fungal disease alternaria leaf spot caused by Alternaria brassicae is a serious problem in cultivation of this crop. Nonavailability of resistance genes within crossable germplasms of Brassica necessitates use of genetic engineering strategies to develop genetic resistance against this pathogen. The pathogenesis related (PR) proteins are group of plant proteins that are toxic to invading fungal pathogens, but are present in plant in trace amount. Thus, overexpression of PR proteins leads to increased resistance to pathogenic fungi in several crops. The PR protein glucanase hydrolyzes a major cell-wall component, glucan, of pathogenic fungi and acts as a plant defense barrier. We report the expression of a class I basic glucanase gene, under the control of CaMV 35S promoter, in Indian mustard and its genetic resistance against alternaria leaf spot. Southern and Northern hybridization confirmed stable integration and expression of the glucanase gene in mustard transgenics. Several independent transgenics were screened in vitro and under poly house conditions for their resistance against Alternaria brassicae. In an in vitro antifungal assay, transgenics arrested hyphal growth of Alternaria brassicae by 15-54%. Under pathogen-challenged conditions in poly house, the transgenics showed restricted number, size and spread of lesions caused by Alternaria brassicae. Also, the onset of disease was delayed in transgenics compared to untransformed parent plants. The results demonstrate potentiality of a PR protein from a heterologous source in developing alternaria leaf spot resistance in Indian mustard.     

Rorippa indica HSPRO2 expression in transgenic Brassica juncea induces tolerance against mustard aphid Lipaphis erysimi

We have previously established that RiHSPRO2, a nematode resistance protein-like homolog from wild crucifer Rorippa indica (L.) Hiern is a potent candidate to control mustard aphid Lipaphis erysimi. The present study further exploits this protein through structure prediction, biosafety assessment and transgenic study. The RiHSPRO2 protein showed an abundance of alpha helices intervened with loops in the homology-based three-dimensional model. No allergenic moiety was found in its amino acid sequence based on homology search in Protein Data Base. The secondary structure of RiHSPRO2 was unstable at temperatures above 50 °C. In vitro pepsin digestion assay revealed the protein to be digested in pepsin supplemented Simulated Gastric Fluid (SGF) within 2 minutes. The protein is proved to be biologically safe as per the FAO/WHO guidelines. An efficient Agrobacterium-mediated Brassica juncea transformation involving direct organogenesis with mean transformation frequency of 5.06?±?0.28% is reported. RiHSPRO2, under the influence of constitutive promoter CaMV35S, was transformed into susceptible B. juncea cv. B85. Southern hybridization confirmed stable integration of the transgene and Western blotting confirmed consistent expression of RiHSPRO2 in the genetically modified Brassica lines. The transgene segregated following Mendelian 3:1 ratio in the successive generation. Detached leaf aphid bioassay and in planta aphid bioassay in transgenic B. juncea lines revealed a reduction in aphid survivability by 45% and a decrease in aphid fecundity by 45.6%.

     

Morphological,ecological and biological variations in the mustard aphid,Lipaphis pseudobrassicae (Kaltenbach) (Hemiptera: Aphididae) from different host plants

Genetic and morphological differentiation of insect populations in relation to the use of different host plants is an important phenomenon that leads to ecological specialization. In this study, we describe variations in morphology, and in ecological and biological parameters of Lipaphis pseudobrassicae (Kaltenbach) clones associated with three host species of Cruciferae, Brassica juncea (L.) var. rai sarson Czern and Cross (brown mustard), Brassica campestris L. var. sarson Prain (yellow mustard), and Rorippa indica (L.) Hiern (wild herb). This study was aimed at obtaining evidence regarding phenotypic differentiation induced by, or associated with, the use of distinct host species. Ten morphological characters, 4 growth parameters and 8 biological functions were investigated in wingless aphids collected from plants of the three host species. Aphids from B. campestris and B. juncea clones were bigger in size, heavier in weight and showed higher growth rates and fecundity than the clones from R. indica. Between the two crop plants, clones from B. juncea showed significantly higher growth rates than the clones from B. campestris. Transfer of L. pseudobrassicae populations from B. campestris to B. juncea and R. indica and vice versa resulted in poor performance. Results indicate that the average phenotype of L. pseudobrassicae individuals inhabiting different host plant species differs as a consequence of the contrasting feeding environments the host species provide.     

Prey-predator relationship between Lipaphis erysimi Kalt. (Hom., Aphididae) and Coccinella septempunctata L. (Col., Coccinellidae). II. Effect of host plants on the functional response of the predator

The present paper examines the effect of the host plants Brassica oleracea, Raphanus sativus and Brassica campestris on which Lipaphis erysimi was reared, on the functional response of the grub of Coccinella septempunctata . The functional response of the grub of C. septempunctata is of type II. The amount of prey consumption increases significantly with the increase of prey density and is maximum in B. oleracea- reared aphids, followed by those reared on R. sativus and B. campestris . The percentage prey consumption declined throughout. This decrease was a maximum for aphids reared on B. campestris followed by R. sativus - and B. oleracea -reared aphids. Analysis of variance also confirmed that the number and percentage prey consumption was significantly influenced between the three host plants and between 12 prey densities. Results show that C. septempunctata has a greater liking for aphids reared on B. oleracea rather than those reared on the other plants that were tested.     

植物提取物和药剂对蔬菜蚜虫实验和自然种群的联合控制作用

研究白蝴蝶（Syngonium podophyllum ）乙醇提取物、苍耳（Xanthium sibiricum ）乙醇提取物、机油乳剂和0.3%印楝素乳油对桃蚜（Myzus persicae）和萝卜蚜（Lipaphis erysimi）的控制效果.室内四因子（1/2实施）二次正交回归旋转组合设计测试对有翅蚜的忌避作用,结果表明,对有翅桃蚜的主要忌避作用物为苍耳乙醇提取物,当苍耳提取物与白蝴蝶乙醇提取物混用,以及机油乳剂和印楝素混用时,对桃蚜有翅蚜的忌避效果提高;而对萝卜蚜有翅蚜的忌避作用主要受苍耳提取物和白蝴蝶提取物的影响.这些干扰作用均是非线性的.田间试验结果,单独使用白蝴蝶提取物对有翅成蚜有较强的驱避作用;对萝卜蚜自然种群的干扰控制作用以4种植物提取物和药剂混配效果最好,达95.7%;对桃蚜自然种群干扰控制效果最好的则是白蝴蝶提取物,控制效果达87%;同时,白蝴蝶提取物与印楝素乳油混配以及苍耳提取物与印楝素乳油混配,对两种蚜虫的控制效果均达80%以上.     

Performance of cabbage stem flea beetle larvae (Psylliodes chrysocephala) in brassicaceous plants and the effect of glucosinolate profiles

The cabbage stem flea beetle (CSFB), Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae), is one of the most important pests in European winter oilseed rape production. Adult beetles feed on young leaves whereas larvae mine within the petioles and stems. Larval infestation can cause significant crop damage. In this study, the host quality for CSFB of four oilseed rape (Brassica napus L.) cultivars and seven other brassicaceous species with different glucosinolate (GSL) profiles was assessed under controlled conditions. Larval instar weights and mortality were measured after 14 and 21 days of feeding in the petioles of test plants. To study the impact of GSL on the performance of larvae, the GSL contents in petioles from non-infested and infested plants were analysed before, and 21 days after, the start of larval infestation. Larval performance was not significantly different between the four cultivars of oilseed rape, but differed considerably among the other brassicaceous species tested. In comparison to the weight of larvae in the standard B. napus cv. Robust, the larval weight was higher in turnip rape (Brassica rapa L. var. silvestris) and significantly reduced in white mustard (Sinapis alba L.), oil radish (Raphanus sativa L. var. oleiformis), and cabbage (Brassica oleracea L. convar. capitata var. alba). The duration of larval development increased in white mustard and oilseed radish. The GSL profiles of the petioles showed little difference between non-infested and infested plants of oilseed rape whereas the content of aliphatic GSL increased in the infested turnip rape plants. In contrast, the aliphatic and benzenic GSL decreased in infested Indian rape (B. rapa subsp. dichotoma Roxb.). Larval weight was not correlated with the total GSL content of plants, neither before infestation nor 21 days after. Larval weight was positively correlated with progoitrin and 4-hydroxyglucobrassicin. White mustard, which provides inferior host quality for larval development, has the potential to introduce insect resistance into high-yielding oilseed rape cultivars in breeding programmes.     

白蝴蝶挥发油对蚜虫的驱避作用及其化学成分

室内用四臂嗅觉仪测试白蝴蝶(Syngonium podophyllum)挥发油对红桃蚜(Myzus persicae)、萝卜(Lipaphis erysimi)和瓜蚜(A[jos gossypii)无翅蚜行为选择的干扰作用．结果表明,在所测试的浓度下,白蝴蝶挥发油对瓜蚜和萝卜蚜有显著的驱避效果,与对照比较,供试蚜虫较少选择处理区,在处理区的停留时间比对照明显缩短;所测试的浓度对红桃蚜无驱避作用．进一步采用气相色谱质谱联用技术对白蝴蝶挥发油进行了成份分析．表明该挥发油小分子主要成份是羰基化合物、脂肪酸和烃．     

Effects of crop diversification levels and fertilization regimes on abundance of Brevicoryne brassicae (L.) and its parasitization by Diaeretiella rapae (M’Intosh) in broccoli

1 The effects of intercropping via competition on crop yields, pest [cabbage aphid Brevicoryne brassicae (L.)] abundance, and natural enemy efficacy were studied in the Brassica oleracea L. var. italica system. 2 From May to December 2004, insect populations and yield parameters were monitored in summer and autumn in broccoli monoculture and polyculture systems with or without competition from Brassica spp. (mustard), or Fagopyrum esculentum Moench (buckwheat), with addition of organic (compost) or synthetic fertilizer. 3 Competition from buckwheat and mustard intercrops did not influence pest density on broccoli; rather, aphid pressure decreased and natural enemies of cabbage aphid were enhanced in intercropping treatments, but this varied with the intercropped plant and season (summer vs. autumn). 4 In compost‐fertilized broccoli systems, seasonal parasitization rates of B. brassicae by Diaeretiella rapae (M’Intosh) increased along with the expected lower aphid pressure compared with synthetically fertilized plants.     

Comparison of feeding behaviour of two Brassica pests Brevicoryne brassicae and Myzus persicae on wild and cultivated brassica species

Feeding behaviour of the specialist Brassicae aphid, Brevicoryne brassicae (L) (cabbage aphid) and the generalist, Myzus persicae, (Sulzer) (peach potato aphid) was monitored electronically on the susceptible cauliflower, Brassica oleracea var. botrytis cv Newton Seale, and a range of 17 Brassica species, B. carinata, B. juncea, B. nigra, B. macrocarpa, and B. villosa var. drepanensis and cultivated brassica varieties, B. oleracea, B. campestris and B. napus. Aphids, monitored for 10 h on the underside of leaves, performed recognisable feeding behaviour on all brassica species. The main differences in feeding behaviour, between M. persicae and B. brassicae, on the susceptible cauliflower Newton Seale, were fewer probes, shorter times to initially reach the phloem but longer times to establish sustained phloem ingestion and the longer times spent, by M. persicae, in xylem ingestion.Feeding behaviour on the range of brassica species tested indicated that generalist and specialist aphids are influenced differently by the host plant. A longer time spent in xylem ingestion was again the major difference in the feeding behaviour of the two aphids. In addition, rejection of passive phloem ingestion, by M. persicae, was not related so closely to increased time spent in non probing activities, as for B. brassicae. This observation indicates that M. persicae does not generally accept or reject brassica species due to the presence of phagostimulants, such as glucosinolates at the leaf surface or along the stylet pathway, unless the concentration is very high. Differences in feeding strategies employed by generalist and specialist aphids on the same plants are discussed.