Identification of the bright-greenish-yellow-fluorescence (BGY-F) compound on cotton lint associated with aflatoxin contamination in cottonseed

In order to characterize the structure of the bright-greenish-yellow-fluorescence (BGY-F) compound on cotton lint associated with aflatoxin contamination in cotton seed, various in vitro and in vivo natural BGY-F reaction products were prepared. Under similar high pressure liquid chromatography separation with variable wavelength and programmable fluorescence detection (HPLC-UV/FL), combined with atmospheric pressure ionization and mass spectral determinations it was found that the BGY-F reaction products prepared from three preparations: (a) kojic acid (KA) + peroxidase (soybean peroxide or horseradish type VI and type II) + H2O2, or (b) detached fresh cotton locules + KA + H2O2, or (c) attached field cotton locules that were treated with a spore suspension of aflatoxigenic Aspergillus flavus, all resulted in identical chromatographic characteristics, and all exhibited a molecular weight of 282. Further characterization of the BGY-F reaction product with 1H- and 13C-NMR spectroscopic analysis revealed that it was a dehydrogenator dimer of 2 KA, linked through the C-6 positions.     

转基因抗草甘膦抗虫棉对棉田冠层节肢动物群落的影响

【背景】随着转基因抗虫棉在我国的广泛种植,一种具有抗虫和耐除草剂（草甘膦）的双价棉被培育成功。这种转双价基因棉和转单价基因抗虫棉对棉田节肢动物群落结构的影响可能不同。因此,在该类转双价基因棉花进行环境释放之前,有必要研究其对棉田节肢动物群落的影响,评价其环境安全性。【方法】试验于2010年5月9日~9月23日和2011年5月10日~9月24日在河南省安阳市中国农业科学院棉花研究所试验农场进行,棉田类型有3种——转基因抗草甘膦抗虫棉田、转Bt棉田和常规棉田,每种棉田种植3个小区,每个小区面积为200 m2（8 m×25 m）。采用对角线5点取样方法,每5 d调查1次棉田的节肢动物群落,通过目测对节肢动物鉴定到属。【结果】抗草甘膦抗虫棉田、Bt棉田和常规棉田节肢动物群落、害虫亚群落和天敌亚群落的结构与组成无明显差异;抗草甘膦抗虫棉田害虫种群数量低于抗虫棉田和常规棉田,而其天敌种群数量与常规棉田相当,略低于Bt棉田;3种棉田节肢动物群落、害虫亚群落和天敌亚群落的多样性指数、均匀性指数均无明显差异。【结论与意义】种植转基因抗草甘膦抗虫棉花不会对棉田节肢动物群落组成造成显著影响。本研究为转基因抗草甘膦抗虫棉田的环境安全性评价提供了依据。     

转Cry1Ac+Cry2Ab棉对棉铃虫的控制作用及对天敌捕食烟粉虱功能反应的影响

文章以转Cry1Ac基因棉(中棉所41)和常规棉(中棉所49)为对照,研究了转Cry1Ac+Cry2Ab基因棉(639020)在棉花生长的关键时期——蕾期(二代棉铃虫发生期)、花期(三代棉铃虫发生期)和花铃期(四代棉铃虫发生期)对棉铃虫的控制作用,同时研究了639020棉田主要捕食性天敌(中华草蛉幼虫、龟纹瓢虫、小花蝽和草间小黑蛛)对烟粉虱的捕食功能,明确了639020棉花在生长的关键时期对棉铃虫的控制效果及对棉田主要捕食性天敌捕食功能反应的影响。结果表明,639020棉花对二代和三代棉铃虫具有良好的控制作用,抗虫性分别比中棉所41提高了52.85%和16.22%,其中前者差异达显著水平,后者差异不显著。在棉花蕾期、花期和花铃期,639020棉田棉铃虫落卵量都比中棉所41棉田和中棉所49棉田低(除二代棉铃虫发生期);棉铃虫幼虫数量都极显著低于常规棉,且都低于防治指标,但与中棉所41棉田无显著差异。639020棉田中华草蛉、龟纹瓢虫、小花蝽和草间小黑蛛对烟粉虱的捕食功能与中棉所41棉田和常规棉田相比无显著变化。研究结果以期为新型转基因棉花环境安全性研究及其外源基因的抗虫遗传效应和生产应用前景进行安全性评价。     

转RRM2基因棉生长势和产量及对棉田节肢动物群落的影响

为了评估新型转基因棉花的育种价值, 评价其对棉田生态环境安全性的影响, 2012-2013年连续2年以新型转RRM2基因棉花(Gossypium hirsutum)为材料, 以其亲本‘中棉所12’为对照, 系统研究了棉田节肢动物群落的结构与组成、群落特征参数及其季节性动态变化, 同时结合气候条件分析比较了转基因棉花和非转基因棉花的生长势和产量构成因素。结果表明: 相同年份新型转RRM2基因棉田昆虫群落、害虫亚群落和天敌亚群落的结构与组成、多样性指数、均匀度指数、优势集中性指数及棉花生长期相对丰度动态变化均与对照无显著差异; 与2012年相比, 2013年转基因棉田和非转基因棉田节肢动物群落、棉花生长及产量构成因素的各项指标总体上呈下降趋势; 转基因棉田和非转基因棉田优势天敌功能团动态均滞后于优势害虫功能团, 因此棉花生长前期应适时防治棉花害虫。     

棉田捕食性瓢虫控害功能的分析

在系统调查棉田捕食性飘虫种数量动态的基础上,应用生态能学的方法,分析并比较了不同插种期、套间作等农业措施对棉田捕食性飘虫控害虫功能的影响。结果表明,不同类型棉田飘虫对害虫的摄入量为26．45－70kJ.m^-2,分别占整个捕食性在天敌摄入量的28．30％－47．88％,是棉田捕食性天敌的优势种类;不同类型棉田飘虫对棉蚜的控害系数为5．07％－12．85％,并随着棉花播种期的推后和间套作呈现出下降趋势,它们在棉田生态系统害虫生态调控中发挥着重要作用。     

Growth vigour and yield of transgenic RRM2 (RNA recognition motif 2) cotton and their effects on arthropod community in cotton field

Aims This experiment was carried out to assess the breeding value of the transgenic RRM2 (RNA recognition motif 2) cotton and to evaluate its environmental safety on cotton field.
Methods The indices of structure and composition, community characteristics, biodiversity, species richness and the seasonal changes of the arthropod community in transgenic RRM2 cotton field and the parental conventional cotton field were studied systematically in 2012 and 2013, respectively. We also analyzed the climate data from June to September of the two years, and compared the cotton growth vigour and the yield parameters between the transgenic cotton and the non-transgenic cotton.
Important findings The results showed that there were no significant differences in the indices of structure and composition, biodiversity, evenness, dominant concentration and the seasonal changes of insect communities and sub-communities between the transgenic cotton field and the non-transgenic cotton field within the same years. Comparing with in 2012, all the indices of arthropod communities and cotton growth vigour and cotton yield parameters in 2013 showed an overall downward trend in both the transgenic and the non-transgenic cotton fields. The dynamics of functional groups of dominant natural enemies lagged behind that of the dominant pests both in the transgenic cotton field and the non-transgenic cotton field, indicating a need to control the pests at the early stage of cotton growth.     

Biological control of cotton pests in China

Cotton is one of the most economically important crops in China, while insect pest damage is the major restriction factor for cotton production. The strategy of integrated pest management (IPM), in which biological control plays an important role, has been widely applied. Nearly 500 species of natural enemies have been reported in cotton systems in China, but few species have been examined closely. Seventy-six species, belonging to 53 genera, of major arthropod predators and parasitoids of lepidoptera pests, and 46 species, belonging to 29 genera, of natural enemies of sucking pests have been described. In addition, microsporidia, fungi, bacteria and viruses are also important natural enemies of cotton pests. Trichogramma spp., Microplitis mediator, Amblyseius cucumeris, Bacillus thuringiensis and Helicoverpa armigera nuclear polyhedrosis virus (HaNPV) have been mass reared or commercially produced and used in China. IPM strategies for cotton pests comprising of cultural, biological, physical and chemical controls have been developed and implemented in the Yellow River Region (YRR), Changjiang River Region (CRR) and Northwestern Region (NR) of China over the past several decades. In recent years, Bt cotton has been widely planted for selectively combating cotton bollworm, H. armigera, pink bollworm, Pectinophora gossypiella, and other lepidopteran pest species. As a result of reduced insecticide sprays, increased abundance of natural enemies in Bt cotton fields efficiently prevents outbreaks of other pests such as cotton aphids. In contrast, populations of mirid plant bugs have increased dramatically due to a reduction in the number of foliar insecticide applications for control of the bollworms in Bt cotton, and now pose a key problem in cotton production. In response to this new pest issue in cotton production, control strategies including biological control measures are being developed in China.     

Screening of abiotic stress‐responsive cotton genes using a cotton full‐length cDNA overexpressing Arabidopsis library

Cotton (Gossypium hirsutum L.) is a major crop and the main source of natural fiber worldwide. Because various abiotic and biotic stresses strongly influence cotton fiber yield and quality, improved stress resistance of this crop plant is urgently needed. In this study, we used Gateway technology to construct a normalized full‐length cDNA overexpressing (FOX) library from upland cotton cultivar ZM12 under various stress conditions. The library was transformed into Arabidopsis to produce a cotton‐FOX‐Arabidopsis library. Screening of this library yielded 6,830 transgenic Arabidopsis lines, of which 757 were selected for sequencing to ultimately obtain 659 cotton ESTs. GO and KEGG analyses mapped most of the cotton ESTs to plant biological process, cellular component, and molecular function categories. Next, 156 potential stress‐responsive cotton genes were identified from the cotton‐FOX‐Arabidopsis library under drought, salt, ABA, and other stress conditions. Four stress‐related genes identified from the library, designated as GhCAS, GhAPX, GhSDH, and GhPOD, were cloned from cotton complementary DNA, and their expression patterns under stress were analyzed. Phenotypic experiments indicated that overexpression of these cotton genes in Arabidopsis affected the response to abiotic stress. The method developed in this study lays a foundation for high‐throughput cloning and rapid identification of cotton functional genes.     

Assessment of cotton aphids, Aphis gossypii, and their natural enemies on aphid-resistant and aphid-susceptible wheat varieties in a wheat–cotton relay intercropping system

The cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae), is an important cotton pest in northern China, especially in the seedling stage of cotton. After large scale commercial use of transgenic Bt cotton, cotton aphids became one of the most important cotton pests. A 2‐year study was conducted to evaluate the role of four winter wheat varieties that were resistant or susceptible to wheat aphid, Sitobion avenae Fabricius (Homoptera: Aphididae), in conserving arthropod natural enemies and suppressing cotton aphids in a wheat–cotton relay intercropping system in northern China. The results indicated that wheat–cotton intercropping preserved and augmented natural enemies more than a monoculture of cotton. The density of natural enemies in cotton was significantly different among relay‐intercropping fields with different wheat varieties. The highest density of natural enemies and low cotton aphid populations were found in the treatment of cotton in relay intercropped with the wheat variety Lovrin10, which is susceptible to wheat aphid. The lowest density of predators and parasitoids associated with high cotton aphid populations were found with the wheat variety KOK1679, which is resistant to wheat aphid. The results showed that wheat varieties that are susceptible or moderately resistant to wheat aphid might reduce cotton aphids more effectively than an aphid‐resistant variety in the intercropping system by enhancing predators to suppress cotton aphids during the cotton seedling stage.     

Simple sequence repeat marker associated with a natural leaf defoliation trait in tetraploid cotton

Cotton (Gossypium hirsutum L.) leaf defoliation has a significant ecological and economical impact on cotton production. Thus the utilization of a natural leaf defoliation trait, which exists in wild diploid cotton species, in the development of tetraploid cultivated cotton will not only be cost effective, but will also facilitate production of very high-grade fiber. The primary goal of our research was to tag loci associated with natural leaf defoliation using microsatellite markers in Upland cotton. The F2 populations developed from reciprocal crosses between the two parental cotton lines--AN-Boyovut-2 (2n = 52), a late leaf defoliating type, and Listopad Beliy (2n = 52), a naturally early leaf defoliating type--demonstrated that the naturally early leaf defoliation trait has heritability values of 0.74 and 0.84 in the reciprocal F2 population. The observed phenotypic segregation difference in reciprocal crosses suggested a minor cytoplasmic effect in the phenotypic expression of the naturally early leaf defoliation trait. Results from the Kruskal-Wallis (KW) nonparametric test revealed that JESPR-13 (KW = 6.17), JESPR-153 (KW = 9.97), and JESPR-178 (KW = 13.45) Simple sequence repeat (SSR) markers are significantly associated with natural leaf defoliation in the mapping population having stable estimates at empirically obtained critical thresholds (P < .05-.0001). JESPR-178 revealed the highest estimates (P < .0001) for association with the natural leaf defoliation trait, exceeding maximum empirical threshold values. JESPR-178 was assigned to the short arm of chromosome 18, suggesting indirectly that genes associated with natural leaf defoliation might be located on this chromosome. This microsatellite marker may have the potential for use to introgress the naturally early leaf defoliation quantitative trait loci (QTL) from the donor line Listopad Beliy to commercial varieties of cotton through marker-assisted selection programs.     

基于棉花功能叶高光谱参数的土壤电导率监测模拟

通过2008-2009年在江苏南京农业大学牌楼试验站的盆栽试验,选择耐盐棉花品种中棉所44和盐敏感性品种苏棉12号为材料,模拟5种不同含盐水平的滨海盐土(0、0.35％、0.60％、0.85％和1.00％),分析了棉花生育期棉田土壤电导率与棉花功能叶光谱反射率和高光谱参数的关系,并建立了棉田土壤电导率(EC)的定量监测模型.结果表明:棉花功能叶光谱反射率在近红外和中红外区域均随土壤盐分水平的升高而升高;以敏感波段1350nm和2307 nm构建的归一化光谱指数NDSI(R1350,R2307)与土壤电导率的决定系数最高,基于此构建了基于NDSI(R1350,R2307)的棉田土壤EC监测模型:EC=-42.899NDSI(R1350,R2307)+27.338;在光谱微分参数中,以TM影像第5个波段的光谱反射率(TM5-SWIR)与棉田土壤EC的决定系数最高,构建了基于TM5-SWIR的棉田土壤EC监测模型:EC=0.0574 TM5-SWIR2-2.5928 TM5-SWIR+ 30.021.以NDSI(R1350,R2307)和TM5-SWIR为自变量的监测模型的预测精度均较高,分别为0.887和0.814,根均方差均较小,分别为1.09和1.29 dS·m-1.利用棉花功能叶NDSI(R1350,R2307)和TM5-SWIR均能较好地监测棉田土壤电导率.     

转RRM2基因棉田昆虫群落多样性及其时序动态

为了评价新型转基因棉花在长江生态区对棉田生态环境安全性的影响, 2013-2015年作者以新型转RRM2基因棉花(Gossypium hirsutum)为材料, 以其亲本‘中棉所12’为对照, 在安徽省沿江棉区系统研究了转基因棉田昆虫群落、害虫亚群落和天敌亚群落的结构与组成、个体数量、群落特征参数及其季节性动态变化。结果表明: 转RRM2基因棉田的主要类群组成、优势类群与非转基因棉田没有差异, 但在2013年转RRM2基因棉田棉蚜个体数量显著高于非转基因棉田, 叶螨类、棉铃虫和其他鳞翅目的个体数量显著低于非转基因棉田, 在其他年份没有显著差异; 其他类群的个体数量在两类棉田间没有显著差异。在棉田害虫发生量大的年份, 转RRM2基因棉田的昆虫群落个体数量较非转基因棉田增加, 物种丰富度较非转基因棉田减少, 但差异不显著, 而两类棉田年度间均差异显著。转RRM2基因棉田昆虫群落和害虫亚群落的全生育期多样性指数、均匀度指数和优势集中性指数与非转基因棉田没有显著差异; 其天敌亚群落的三个指数在2013年与非转基因棉田差异显著, 其他年份没有显著差异; 两类棉田年度间差异均不显著。转RRM2基因棉田昆虫群落、害虫亚群落和天敌亚群落个体数量、群落特征参数的时序动态与非转基因棉田较一致, 具季节性波动; 在群落个体数量高峰期, 群落多样性指数和均匀度指数处于低谷, 而优势集中性指数则相反; 昆虫群落、害虫亚群落的季节波动明显, 天敌亚群落的季节性变化较平缓。因此, 转RRM2基因棉花对棉田昆虫群落的结构与组成、群落特征参数及其时序动态没有显著影响, 但在气候适宜年份, 转RRM2基因棉田的害虫发生量可能增大。     

Influence of light and habitat on predation of Culex quinquefasciatus （Diptera： Culicidae）larvae by the waterbugs （Hemiptera： Heteroptera）

The influence of light and habitat structure on the predation of Culex quinquefasciatus larvae by the common heteropteran water bug, Diplonychus （= Sphaerodema） annulatus, D. rusticus and Anisops bouvieri was assessed in the laboratory. It was revealed that water bugs predate more in presence of light than in dark conditions. While A. bouvieri consumed more prey in structured conditions, D. annulatus and D. rusticus consumed more prey in open conditions. The selection of prey size as well as the respective numbers varied between predators and treatments significantly. Prey vulnerability （PV）, an indicator of predatory efficiency, was highest for D. annulatus, moderate for D. rusticus and low for A. bouvieri. Prey consumption and PV values under different treatment conditions indicate that for belostomatid water bugs D. annulatus and D. rusticus, the order of prey consumption under different habitat and light/dark combinations is light open 〉 dark open 〉 light vegetated 〉 dark vegetated. In the case of the backswimmer, A. bouvieri, the order of prey consumption appeared to be light structured 〉 dark structured 〉 light open 〉 dark open. These findings were consistent with resource-partitioning by water bug species sharing the same guild. If the observed results are extended to natural settings the efficiency of these predators in controlling mosquito populations will vary with the structural complexity of habitats and the intensity of light.     

BT基因棉与常规棉主要害虫及天敌生态位的比较研究

本研究了BT基因棉与常规棉主要害虫及天敌的生态位。结果表明：主要害虫及天敌的种类一致,但棉铃虫幼虫的数量差异显;33B棉田各昆虫种群的生态位宽度指数偏高;棉铃虫、棉蚜对主要天敌的生态位重叠指数偏高。应用生态位概念分析了造成区别的原因。     

Impact of transgenic cotton plants on a non-target pest, Aphis gossypii Glover

Abstract.  1. Transgenic crops have shown great promise for the control of target pest insects, but in some cases they can also influence non-target species. This study investigated the impact of Bt and Bt+CpTI transgenic cottons on the non-target cotton aphid, Aphis gossypii Glover, by comparing life-table parameters, feeding behaviour, and the fluctuating asymmetry of morphological traits of aphids reared on transgenic cotton and those on untransformed control plants.
2. Aphids on the Bt+CpTI cotton showed a shorter reproductive duration and maximum lifespan, lower survival rates and potential maximum fecundity, and an earlier occurrence of peak daily mortality in the first or second generation. However, the aphid population soon developed fitness and overcame the negative effect in the second or third generation. The aphids on the Bt cotton had significantly longer reproductive durations in the first generation, higher survival rates in the third generation, and apparently larger potential maximum fecundity in all three generations.
3. The percentages of accumulated duration of feeding waveforms E1 and E2 were significantly lower in aphids on the Bt+CpTI cotton than in those on the Bt or control cotton, whereas the frequencies of moving and finding feeding sites, and probe behaviour were significantly higher.
4. Fluctuating asymmetry in three morphological characters of aphids reared on transgenic and control cotton was detected. The fluctuating asymmetry value of the third segment of antenna in aphids on Bt+CpTI cotton was significantly higher than that of aphids on Bt or control cotton. Based on the fluctuating asymmetry value, the stress of cotton on the aphids could be ranked as Bt+CpTI cotton > Bt cotton > control cotton.     

Isolation and characterization of insecticidal activity of （Z）- asarone from Acorus calamus L.

The insecticidal activity of Acorus calamus L. rhizome-derived material against adults of Sitophilus zeamais Motschulsky was examined by using repellency method and contact toxicity. The biologically active constituent of the A. calamus rhizome was separated and identified. The results showed that the ethanol extract of A. calamus had strong repellency and contact effect to S. zeamais and the active constituent of the A. calamus was characterized as （Z）-asarone by spectroscopic analysis. Responses from the tests varied with exposure times and doses. In the repellency test, ethanol extract of A. calamus had 93.92% repellency at 629.08 μg/cm^2 but only 71.38% at 157.27 μg/cm^2 12 h after treatment. As a contrast, （Z）-asarone showed 84.50% repellency at 314.54μg/cm^2 and 77.02% at 78.63 μg/cm^2 12 h after treatment. In the filter paper diffusion test, ethanol extract of A. calamus caused 95.56% and 17.78% mortality to S. zeamais at 314.54 μg/cm^2 and 78.63 μg/ cm^2 4 days after treatment, while （Z）-asarone brought about 100.00% and 15.56% mortality at 40.89 μg/cm^2 and 15.73 μg/cm^2 respectively. These results indicate that the insecticidal activity of the A. calamus extract may be due to （Z）-asarone.     

Proteomics profiling of fiber development and domestication in upland cotton (Gossypium hirsutum L.)

Comparative proteomic analyses were performed to detail the evolutionary consequences of strong directional selection for enhanced fiber traits in modern upland cotton (Gossypium hirsutum L.). Using two complementary proteomic approaches, 2-DE and iTRAQ LC–MS/MS, fiber proteomes were examined for four representative stages of fiber development. Approximately 1,000 protein features were characterized using each strategy, collectively resulting in the identification and functional categorization of 1,223 proteins. Unequal contributions of homoeologous proteins were detected for over a third of the fiber proteome, but overall expression was balanced with respect to the genome-of-origin in the allopolyploid G. hirsutum. About 30 % of the proteins were differentially expressed during fiber development within wild and domesticated cotton. Notably, domestication was accompanied by a doubling of protein developmental dynamics for the period between 10 and 20 days following pollination. Expression levels of 240 iTRAQ proteins and 293 2-DE spots were altered by domestication, collectively representing multiple cellular and metabolic processes, including metabolism, energy, protein synthesis and destination, defense and stress response. Analyses of homoeolog-specific expression indicate that duplicated gene products in cotton fibers can be differently regulated in response to selection. These results demonstrate the power of proteomics for the analysis of crop domestication and phenotypic evolution.     

绿盲蝽对不同生长期棉花的刺吸危害特性

【目的】近年绿盲蝽Apolygus lucorum (Meyer-Dür)对棉花的危害成为农业中热议的话题和重点研究的课题。为更加深入了解此种害虫对棉花各组织的危害特性及量化危害,为早期预测预报提供依据,我们研究了绿盲蝽对棉花不同生长阶段（各组织）早期危害表征与后期成铃情况的关系。【方法】通过大田接虫法研究了绿盲蝽对苗期、蕾期、花期和铃期棉花植株的危害特性。【结果】绿盲蝽危害24 h会造成棉花子叶期和2叶期产生无头苗,2叶期后受绿盲蝽危害不会形成无头苗。建立了无头苗率和生长点被害级别的关系模型y=0.1906x-0.0439 (1≤x≤5)和棉花苗期（不同叶期）被害10 d后的叶片被害指数与刺吸点数和生长点被害级别回归模型：4叶期：y=0.532+0.202x₁+0.005x₂; 6叶期：y=-0.063+0.339x₁+0.002x₂; 8叶期：y=-0.150+0.087x₁+0.001x₂ (0≤x₁≤5, 1≤x₂≤5)。绿盲蝽对蕾和花的刺吸危害没有对后期棉铃的脱落率和大小产生显著影响。10 mm和15 mm直径棉铃被害后的校正脱落率分别为55.00%和26.60%,而直径大于15 mm的棉铃校正脱落率均为0;绿盲蝽的刺吸危害对棉铃的成铃大小无显著影响;建立了幼铃（10 mm、10~15 mm直径棉铃）刺吸点数和脱落率的回归模型［10 mm: y=0.1857x-0.081; 10~15 mm: y=0.0522x-0.0068（1≤x≤5）］。【结论】结果表明,幼苗阶段,棉花的子叶期和2叶期是受绿盲蝽危害的敏感时期;成株阶段,棉花嫩铃期是受绿盲蝽危害的敏感时期;建立的绿盲蝽刺吸危害的回归模型可用于绿盲蝽危害损失短期预测。因此,应在棉花受害敏感时期对绿盲蝽重点防治,提高防治效率。