Pink bollworm moth （Lepidoptera： Gelechiidae） catches in the Imperial Valley, California from 1989 to 2003

We examined the patterns of male pink bollworm （PBW）, Pectinophora gossypiella （Saunders）, moth catches in gossyplure-baited traps over a 15-year period from 1989 to 2003 in the Imperial Valley, California, USA. Monitoring was conducted during periods when different pink bollworm areawide control strategies were being used. Numbers of male pink bollworm moths caught in gossyplure-baited traps progressively decreased each year from 1990 to 1994 during short-season cotton production. High numbers of male moths caught in traps from 1995 to 1997 may have been related to moth migrations from the large cotton acreages grown in the Mexicali Valley bordering the Imperial Valley. Transgenic Bollgard （Bt） cotton was planted in 3% of the cotton area in 1996 and thereafter in 80%- 94% of the cotton area from 1997 to 2003. Pink bollworm moth trap catches were significantly lower from 1998 to 2003 than catches in 1995 to 1997, except for 1999. The trapping results suggested that Bt cotton had significant input on reduction of pink bollworm populations, confirming results of other investigators and providing additional documentation on the benefits of the Bt cotton culture.     

BIOLOGY OF CHRYSOPA PHYLLOCHROMA WESMAEL （NEUROPTERA： CHRYSOPIDAE）. Ⅱ： INTRASPECIFIC INTERFERENCE AND SEARCHING CAPACITY

The present study examined intraspecific interference and searching behavior of Chrysopa phyllochroma Wesmael (Neuroptera: Chrysopidae) for Aphis gossypii Glover (Homoptera: Aphididae) nymphs under laboratory and greenhouse conditions. The results were shown as follow: 1) In four different arenas (i.e. Petri dish, glass vessel, glass vessel with barriers in it, and cage with potted cotton plant), the predaceous efficiency of C. phyllochroma larvae varied with the predator density, the hunt constant (Q) and the intraspecific interference (m)increased with the prey density but decreased with the space heterogeneity; 2) In cage with potted cotton plant, the first- and second-instar green lacewing larvae consumed 13.6 and 29.4 cotton aphids/day respectively. The number of cotton aphids consumed by C. phyllochroma on lower leaves was significantly less than that on upper leaves; and 3) In cage with potted cotton plant, the percentage of the first- and second-instar green lacewing larvae located on upper leaves was significant less than that on lower leaves.     

Preliminary studies on differential defense responses induced during plant communication

We compared the expression patterns of three representative genes in undamaged tomato and tobacco plants in response to exposure to either tomato or tobacco fed on by Helicoverpa armigera (cotton bollworm). When tomato and tobacco, two species of one family, were incubated in the chambers with the tomato plants damaged by the cotton bollworm, the expression of the PR1, BGL2, and PAL genes was up-regulated in leaves of both plants. However, the levels of gene expression were significantly higher in the tomato than that in the tobacco. In addition, the activities of enzymes, peroxidase, polyphenol oxidase, and lipoxygenase were found to be higher in the tomato than those in the tobacco. Similar results were obtained when the damaged plants were replaced by the tobacco.     

Effects of elevated CO2 on the foraging behavior of cotton bollworm, Helicoverpa armigera

Effects of elevated CO2 on the foraging behavior of cotton bollworm Helicoverpa arrnigera Hübner reared on milky grains of spring wheat grown in ambient, 550μL/L and 750μL/L CO2 concentration atmospheres in open-top chambers （OTC） were studied. The results indicated that： （i） elevated CO2 significantly affected both the type and amount of food eaten by H.arrnigera reared on milky grains of ambient CO2-grown wheat were significant higher than those for bollworm larvae reared on wheat grains grown in 550 and 750μL/L CO2 atmospheres; （ii） when bollworm larvae were reared on mixed milky grains from different CO2-grown wheat （food-choice condition）, larval duration increased significantly-pupal weight, adult longevity, and fecundity decreased significantly, comparing with those reared on milky grains of ambient CO2-grown wheat, 550μL/L CO2-grown wheat and 750μL/L CO2-grown wheat respectively; （iii） significant decreases in the contents of fructose and gross protein （GP） and significant increases in the contents of glucose, amylose, total saccharides （TSC）, TSC： GP ratio, free amino acids and soluble protein in the wheat grains with CO2 rising; （iv） and selected-foraging amount/food-choice index of cotton bollworm H.armigera were significantly positive correlated with the contents of fructose and GP of wheat grains, but they had significantly negative relationships with the contents of glucose, amylose, TSC and TSC： GP ratio of wheat grains.     

Chromosomal deletions mediated by CRISPR/Cas9 in Helicoverpa armigera

Helicoverpa armigera, cotton bollworm, is one of the most disastrous pests worldwide, threatening various food and economic crops. Functional genomic tools may provide efficient approaches for its management. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, dependent on a single guide RNA (sgRNA), has been used to induce indels for targeted mutagenesis in cotton bollworm. However, genomic deletions may be more desirable to disrupt the function of noncoding genes or regulatory sequences. By injecting two sgRNAs with Cas9 protein targeting different exons, we obtained predictable genomic deletions of several hundred bases. We achieved this type of modification with different combinations of sgRNA pairs, including HaCad and HaABCC2. Our finding indicated that CRISPR/Cas9 can be used as an efficient tool to engineer genomes with chromosomal deletion in H. armigera.     

How does the host-specialized aphid deal with food deficiency？

Aphis gossypii Glover shows obvious host specialization, with cucurbit- and cotton-specialized biotypes or host races in many regions. Because its annual natal hostcrops senesce earlier the cucurbit-specialized biotype may suffer food deficiency. The method this biotype uses to overcome this challenge is still poorly understood. In orderto understand the potential of the cucurbit-specialized biotype aphids in host shift and usage, the performance of this biotype on cotton （Gossypium hirsutum）, a common butpoor quality host plant, was explored in this study. The cucurbit-specialized aphids could establish populations on cotton only when these plants had at least nine leaves, and subsequent populations developed rather slowly. The presence of whitefly populations on cotton improved the success rate of cucurbit-specialized aphids. The cucurbit-specialized aphidswere mainly distributed on the older leaves of cotton, with only a few settling on the upper leaves. The cucurbit-specialized aphids reared on cotton for 40, 54 and 61 days stillmaintained strong preference for their natal host plant, cucumber （Cucumis sativus）, rather than cotton, and their net reproductive rates and intrinsic rates of natural increase weredramatically lower when they were transferred onto new six-leaf cotton plants or detached leaves. Therefore, we concluded that the cucurbit-specialized aphids have the potentialto utilize mature or whitefly-stressed cotton plants, but that this feeding experience on cotton did not alter their specialization for cucurbits. Some cotton plants could act as atemporary host for the cucurbit-specialized aphids to overcome food deficiency arising from senescing cucurbits.     

Phototoxic effect of UVR on wild type, ebony and yellow mutants of Drosophila melanogaster : Life Span, fertility, courtship and biochemical aspects

Melanin plays an important role in protecting organisms from ultraviolet radiation (UVR). Therefore, it is possible that differently colored strains can show different sensitivities to UVR. In the present work, life span, fertility and courtship behavior of wild type (w), ebony (e) and yellow (y) strains of Drosophila melanogaster were studied to evaluate their sensitivity to ultraviolet (UV). Because a range of photo- toxic effects of UVR are mediated through generation of free radicals, levels of free radicals, lipid per- oxide (malondialdehyde, MDA) and superoxide dismutase (SOD) activity of three strains were examined to indicate their antioxidant defending ability and oxidative status. It was shown that w always had the highest lifespan and fertility not only in the control but also in UV-exposed groups. Moreover, lifespan and fertility of e were significantly higher (P<0.0001) than those of y in the UV-exposed groups, but not for the control. On the other hand, UV exposure had an adverse effect on courtship of flies. Stronger electron paramagnetic resonance (EPR) signals could be detected in w, e and y exposed to 5 min UV. And there were more significant changes of EPR signals in y than in w and e. UVR had no significant (P=0.1782) effect on the SOD activities. After pooling data from the control and UV-exposed groups, we found that w had a significantly (P<0.05) higher level of SOD activity, but e and y were nearly at the same levels (P>0.05). MDA levels were increased in the UV dose-dependent manner (P=0.0495). In con- clusion, our results suggested that UVR can decrease life span and fertility of flies and do harm to courtship, which may be due to oxidative damage to flies tissues (e.g. central nervous system) induced by free radicals. w had the highest tolerance to UVR, which may be ascribed to its advantage of survival under the natural condition and at high level of SOD activity. Then differences of pigment between e and y in absorbing UV, shielding against UV and scavenging free radicals produced by UVR should be responsible for their different sensitivity to UVR.     

A SURVEY OF SIZES AND WEIGHTS OF BEMISIA TABACI（HOMOPTERA： ALEYRODIDAE）B BIOTYPE LIFE STAGES FROM FIELD GROWN COTTON AND CANTALOUPES

Size and weight measurements were made for all the life stages of Bemisia tabaci (Gennadius) B biotype from field grown cotton ( Gossypium hirsutum L. ) and cantaloupe ( Cucumis melo L., var. cantalupensis )in Phoenix, AZ and Fargo, ND, USA in 2000 and 2001. Nymphal volumes were derived from the measurements.The average nymphal volume increase for settled 1 st to the late 4th instar was exponential. The greatest increase in body volume occurred during development from the 3rd to early 4th instar. Nymphs on cotton leaves were wider,but not longer compared with those on cantaloupe. Ventral and dorsal depth ratios of nymphal bodies from 1st tolate 4th instars from cantaloupe leaves were significantly greater compared with those from cotton leaves. During nymphal development from 1st to 4th instar, the average (from the two host species) ventral body half volume in-creased by nearly 51 times compared with an increase of 28 times for the dorsal body half volume. Adult female and male average lengths, from heads to wing tips, were 1 126 μm and 953 μm, respectively. Average adult fe-male and male weights were 39 and 17 μg, respectively. Average widths, lengths, and weights of eggs from cottonand cantaloupe were, 99 μm, 197 μm, and 0.8 μg, respectively. Average widths, lengths, and weights for exu-viae of non-parasitized nymphs from both cotton and cantaloupe were 492 μm, 673 μm, and 1.20 μg, respective-ly; and widths, lengths, and weights of parasitized nymph exuviae were 452 μm, 665 μm, and 3.62 μg, respec-tively. Both exuviae from non-parasitized and parasitized nymphs from cotton leaves were wider, longer, and heavier than those from cantaloupe leaves.     

Structural and Functional Damage Caused by Boron Deficiency in Sunflower Leaves

Boron deficiency induced a dramatic inhibition in sunflower plant growth, shown by a reduction in dry mass of roots and shoots of plants grown for 10 d in nutrient solution supplied with 0.02 μM B. This low B supply facilitated the appearance of brown purple pigmentation on the plant leaves over the entire growth period. Compared to B-sufficient (BS) leaves, leakage from B-deficient (BD) leaves was 20 fold higher for potassium, 38 fold for sucrose, and 6 fold for phenolic compounds. High level of membrane peroxidation was detected by measuring peroxidase activities as well as peroxidative products in BD sunflower plants. Soluble and bound peroxidase activities measured in BD thylakoid membranes were accelerated two fold compared to those detected in BS-membranes. No detectable change in soluble peroxidase activity in roots whereas a 4 fold stimulation in bound peroxidase activity was detected. Thylakoid membranes subjected to low B supply showed enhancement in lipoxygenase activity and malondialdehyde (MDA) content in parallel with 40 and 30 % decrease of linoleic and linolenic acid contents (related to total unsaturated fatty acids). A slower rate of Hill reaction activity (40 %) and a suppressed flow of electron transfer of the whole chain (30 %) were detected in BD thylakoid membranes. This reduction was accompanied with a decline in the activity of photosystem 2 shown by a diminished rate of oxygen evolution (42 %) coupled with a quenching (27.5 %) in chlorophyll a fluorescence emission spectra at 685 nm (F₆₈₅). Thus B is an important element for membrane maintenance, protection, and function by minimizing or limiting production of free oxygen radicals in thylakoid membranes of sunflower leaves. This revised version was published online in June 2006 with corrections to the Cover Date.     

Bollworm responses to release of genetically modified Helicoverpa armigera nucleopolyhedroviruses in cotton

Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) has been developed as a commercial biopesticide to control the cotton bollworm, H. armigera, in China. The major limitation to a broader application of this virus has been the relative long time to incapacitate the target insect. Two HaSNPV recombinants with improved insecticidal properties were released in bollworm-infested cotton. One recombinant (HaCXW1) lacked the ecdysteroid UDP-glucosyltransferase (egt) gene and in another recombinant (HaCXW2), an insect-selective scorpion toxin (AaIT) gene replaced the egt gene. In a cotton field situation H. armigera larvae treated with either HaCXW1 or HaCXW2 were killed faster than larvae in HaSNPV-wt treated plots. Second instar H. armigera larvae, which were collected from HaCXW1 and HaCXW2 treated plots and further reared on artificial diet, showed reduced ST(50) values of 15.3 and 26.3%, respectively, as compared to larvae collected from HaSNPV-wt treated plots. The reduction in consumed leaf area of field collected larvae infected with HaCXW1 and HaCXW2 was approximated 50 and 63%, respectively, as compared to HaSNPV-wt infected larvae at 108 h after treatment. These results suggest that in a cotton field situation the recombinants will be more effective control agents of the cotton bollworm than wild-type HaSNPV.     

Field inactivation of wild-type and genetically modified Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus in cotton

Cotton bollworm (Helicoverpa armigera) is a serious pest on cotton in China. A specific baculovirus, H. armigera nucleopolyhedroviruses (HaSNPV) is used as a commercial biopesticide to control this pest. To improve the pesticidal properties, HaSNPV has been genetically engineered by both deleting the ecdysteroid UDP-glucosyltransferase (egt) gene from its genome (recombinant HaSNPV-EGTD) and incorporating an insect-selective toxin gene from the scorpion Androctonus australis (AaIT) (recombinant HaSNPV-AaIT). In the field, there was no significant difference among the inactivation rates of the two recombinant HaSNPVs and their parent wild-type, HaSNPV-WT. The inactivation rate of these viruses was significantly different in different years. The average half-life of HaSNPV was 0.57, 0.90 and 0.39 days in 2000, 2001 and 2002, respectively. Inactivation rates correlated well with solar radiation over these years.     

一种棉铃虫多角体病毒的纯化与鉴定

对一株纯化的棉铃虫多角体病毒进行电镜观察,形态呈不规则型,直径约2 μm,命名为棉铃虫多角体-B(Helicoverpa armigera nucleocapsid nucleopolyhedrovirus-B,HaNPV-B),它对东方粘虫有较好的感染性,与棉铃虫单核型多角体病毒(Helicoverpa armigera single nucleocapsid NPV,HaSNPV)有不同的宿主域.通过半补齐法建立HaNPV-B基因组文库,拼接了一个6 035 bp的核苷酸片段,推测包含6个ORF,其包含基因内容及基因结构与HaSNPV不同,分析表明两者是不同种病毒,这为杆状病毒与宿主的相互作用及共同进化提供了信息.     

Bt棉叶对棉铃虫抗虫性的时空变化及气象因素的影响

用叶片喂饲法,测定了转Bt基因棉不同叶位、不同生长时期的叶片对棉铃虫初孵幼虫抗虫性的时空变化,同时测定了大田和室内不同气象条件下生长的侧枝和苗期叶片对棉铃虫抗虫性的影响。结果表明：转Bt基因棉R₁₉-137株系主茎第2～10叶的抗虫性最高,初孵幼虫处理5天后的平均死亡率为97.0%～100%,为害级别在1.0～1.1级;第11～16叶的抗虫性明显下降,平均死亡率为35.6%～67.6%,存活幼虫以2龄为主。7月下旬、8月下旬测定了不同部位侧枝叶片的抗虫性,平均死亡率分别为30.9%～44.9%和10.0%～30.0%,抗虫效果进一步下降。试验结果还表明,在室内外不同气候条件下生长的Bt棉叶片的抗虫性有显著差异。讨论了Bt棉抗虫性时空变化的可能原因与Bt棉推广中气候条件的重要性。     

小麦植株挥发性次生物质对棉铃虫的行为及EAG反应

本文用无水乙醚及水蒸汽蒸馏两种方法,分别提取小麦花和叶各1 000g和3 200g。4个提取样品分别用氮气浓缩至2 mL。然后用风洞和触角电位检测棉铃虫Helicoverpaarmigera对小麦花、叶提取物的行为及电生理的反应。风洞实验结果表明：小麦花、叶中所含的挥发性次生物质对棉铃虫有很好的引诱作用,其定向率为45%～72%,高于对照22.2%,经‘检验差异显著;此外,雌蛾对花、叶的定向率均高于雄蛾;而花与叶的提取物对雌、雄蛾的定向率之间无显著差异。触角电位实验结果表明：棉铃虫触角对小麦花、叶的提取物均有强的EAG反应,与对照相比有显著性差异;雌虫的EAG反应略高于雄虫,但差异不显著;小麦花提取物对棉铃虫EAG反应略强于小麦叶,但差异不显著。     

人工释放赤眼蜂对棉铃虫的防治作用及相关生态效应

1998～2000年在河北省南皮县棉区转Bt基因棉田及常规棉田中设置4种不同的组合处理,通过释放螟黄赤眼蜂控制棉铃虫的方法,减少化学农药的使用,达到提升和增强自然生物控制力的生态效应;并以不采取任何防治措施的常规棉田为常规棉对照田。研究表明：（1）转基因棉和常规棉棉田中自然寄生率随棉铃虫世代的增加而逐渐升高,2、3和4代棉铃虫卵被寄生率范围分别为13.3%～14.3%、26.7%～28.2%和60.8%～61.4%。（2）棉铃虫2代期,在常规棉综防田释放赤眼蜂2次,其寄生率为46.4%, 比未释放赤眼蜂的转基因棉棉田和常规棉对照田提高33.1%和32.1%; 3代期,转基因棉棉田释放赤眼蜂1次,其寄生率与常规棉综防田释放4次的效果相当,分别为73.7%和68.1%;但与转基因棉不放蜂田、常规棉化防田及对照田相比,分别提高45.5%、61. 8%和47.0%;4代期,无论放蜂与否,各处理棉田中的寄生率除化防田外（52.1%）均在60%以上。（3）常规棉化防田棉铃虫2代和3代期,分别使用农药2和3次,其自然寄生率分别为5.5%和11.9%,与对照田相比,分别降低8.8%和14.8%;与常规棉综防田相比,分别降低40.9%和56.2%;释放赤眼蜂的效果与施药时间有关,放蜂后1天内施药,寄生率仅为12.5%,施药后2天放蜂,寄生率达45.6%。（4）转基因棉棉田棉铃虫累计数和百株蕾铃被害数比常规棉综防田分别减少74.8%和73.8%,捕食性天敌增加63.0%;放蜂Bt棉田比不放蜂Bt棉田棉铃虫累计数、百株蕾铃被害数分别减少61.8%和33.3%;常规棉综防田棉铃虫累计数、百株蕾铃被害数、农药使用量比化防棉田分别减少29.7%、43.4%和60.0%,捕食性天敌数量增加63.0%。（5）转基因棉田和综防棉田的益害比（捕食性天敌/植食性害虫）分别为0.47∶1和0.30∶1, 而化防棉田为0.24∶1。上述结果表明释放赤眼蜂可弥补抗虫棉后期抗性减弱的不足,增加田间自然天敌的数量,提高对棉田害虫的自然控害功能。     

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。     

The halo effect: suppression of pink bollworm on non-bt cotton by bt cotton in china

In some previously reported cases, transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have suppressed insect pests not only in fields planted with such crops, but also regionally on host plants that do not produce Bt toxins. Here we used 16 years of field data to determine if Bt cotton caused this "halo effect" against pink bollworm (Pectinophora gossypiella) in six provinces of the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We found that Bt cotton significantly decreased the population density of pink bollworm on non-Bt cotton, with net decreases of 91% for eggs and 95% for larvae on non-Bt cotton after 11 years of Bt cotton use. Insecticide sprays targeting pink bollworm and cotton bollworm (Helicoverpa armigera) decreased by 69%. Previously reported evidence of the early stages of evolution of pink bollworm resistance to Bt cotton in China has raised concerns that if unchecked, such resistance could eventually diminish or eliminate the benefits of Bt cotton. The results reported here suggest that it might be possible to find a percentage of Bt cotton lower than the current level that causes sufficient regional pest suppression and reduces the risk of resistance.     

Depressed performance and detoxification enzyme activities of Helicoverpa armigera fed with conventional cotton foliage subjected to methyl jasmonate exposure

Methyl jasmonate (MeJA)‐mediated defense in conventional cotton, Gossypium hirsutum L. (Malvaceae), against cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), was investigated with respect to the activities of the detoxification enzymes acetylcholinesterase (AChE), carboxylesterase (CarE), and glutathione S‐transferases (GST) in pupae as well as the performance of larvae. The results suggested that exogenous application of MeJA to cotton leaves depressed the activities of AChE, CarE, and GST of cotton bollworm pupae. Both the absolute and protein‐specific AChE activities of pupae were depressed at all three MeJA concentrations applied as compared with a control, and the effects of 0.4 mM MeJA were significantly higher than those of 0.1 and 0.2 mM. A marked reduction in absolute CarE activity was observed at the 0.4 mM MeJA treatment, whereas the protein‐specific activity was increased by 0.2 and 0.4 mM. Absolute GST activity was significantly depressed only by the 0.4 mM MeJA treatment, whereas protein‐specific GST activity was not markedly affected by MeJA. Protein content of pupae was reduced by 0.4 mM MeJA‐induced defense in cotton leaves. The development time of larvae was protracted and pupal weight was reduced by 0.1 and 0.4 mM MeJA‐treated cotton leaves. Larval weight gain was inhibited significantly on 0.2 and 0.4 mM MeJA‐treated cotton leaves. The results suggested that MeJA‐induced plant defense may have adverse effects on H. armigera. In addition to the inhibition of growth and development, induced defense may also impair the insect's ability to detoxify toxic plant secondary metabolites.