转Bt基因棉挥发性气味的化学成分及其对棉铃虫的电生理活性

报道了对转Bt基因棉“GK-12”及其常规棉亲本“泗棉3号”挥发性化学物质的分析结果,以及棉铃虫对Bt棉挥发性化学物质的电生理反应,以期为抗虫棉的生态学和安全性评价提供化学生态学的证据。结果表明,在7～8个真叶期,转Bt基因棉的α-蒎烯和β-蒎烯的相对含量比常规棉高许多,倍半萜烯C和一个含量很低的化合物（该化合物对棉铃虫有电生理活性）是常规棉所没有的。转基因Bt棉的挥发性物质中有9个化合物可引起棉铃虫的触角电位反应。     

云南切梢小蠹对云南松树干和枝梢挥发性物质趋向反应的差异分析

利用GC-MS联用仪和计算机信息检索法,通过对云南松Pinus yunnanensis树干韧皮部和枝梢的挥发性物质成分的分析发现,二部位所含挥发性物质在组成成分和含量上存在明显差异.云南松树干挥发物以α-蒎烯为主,含量达81.01%,枝梢为挥发物以α-蒎烯和β-水芹烯为主,含量分别达29.20%和30.52%.与云南松...     

9 种海洋硅藻挥发性成分的比较分析

采用顶空固相微萃取气相色谱一质谱联用技术,通过SIMCA．P分析软件,对硅藻门下处于生长平台后期的9种海洋硅藻的挥发性成分进行比较分析研究。结果表明：在平台后期硅藻的挥发性成分中,醛类物质的含量和种类占据了很大的优势,壬醛、2,4-辛二烯醛、2-己烯醛、2-壬烯醛、2,4-庚二烯醛等是硅藻挥发性成分的主要构成物质,十四烷、十五烷、1-十五烯、十六烷等烷烃类是硅藻中含量仅次于醛类物质的挥发性成分,3,5-辛烯基-3-酮等短链酮类和1-辛烯基-3-醇是硅藻中含量较高的酮类和醇类物质,9种硅藻中均检测二甲基硫的存在。差异性较大的挥发性物质主要有壬醛、8-十七碳烯、1-戊烯．3．酮、1-己烯-3．醇、2,6-二甲基一苯酚,它们决定了不同的硅藻有着各自不同的风味。研究结果表明这些挥发性物质对硅藻的饵料价值、养殖生物的肉质风味、水环境的异味形成有着重要意义。     

棉蚜和七星瓢虫对紫茎泽兰挥发物的行为反应及挥发物化学成分初步分析

田间调查 发现紫茎泽兰Eupatorium adenophorum Spreng上有一定数量的棉蚜Aphis gossypii寄生。从棉蚜是否可为紫茎泽兰的生物防治提供新思路的角度出发,研究了紫茎泽兰挥发性气味物质的化学组成及其对棉蚜和七星瓢虫Coccinella septempunctata行为的影响。棉蚜选择性试验结果表明：紫茎泽兰饲养的棉蚜对紫茎泽兰的趋向率为62.5%,对棉花的趋向率为37.5%。棉花饲养的棉蚜对紫茎泽兰的趋向率为30%,对棉花的趋向率为70%。在七星瓢虫对感染蚜虫植株的选择性实验中,七星瓢虫对感染蚜虫的紫茎泽兰植株气味趋向率为29.2%,对感染蚜虫的棉花植株气味趋向率为70.8%;七星瓢虫对除去植株的两种蚜虫气味选择率没有显著差异,对取食紫茎泽兰的蚜虫气味选择率为46.7%,对取食棉花植株的蚜虫气味选择率为53.3%。通过GC-MC分析了紫茎泽兰挥发性化学物质的组分及相对含量,其主要挥发物的成分是α-蒎烯、莰烯、β-蒎烯、2-蒈烯和α-水芹烯等。     

枫香叶采后挥发物表征及影响因子研究

以枫香(Liquidambar formosana)叶为材料,采用气相色谱/质谱(GC/MS)联用技术对不同发育程度、干燥时间、储存方式及萃取温度下枫香叶挥发性成分进行分析,以揭示枫香叶释放挥发性有机化合物数量及不同采后处理下挥发物的消长规律。结果表明：(1)枫香叶释放的萜烯类化合物以α-蒎烯、β-蒎烯、β-水芹烯、柠檬烯为主;(2)嫩叶挥发性成分种类较成熟叶高,莰烯、β-萜品烯、罗勒烯仅存在于嫩叶,3-蒈烯、γ-松油烯仅存在于成熟叶;(3)干燥时间对枫香叶挥发性化合物影响较大,挥发物种类随干燥时间延长呈逐渐下降趋势,但主要的萜烯类化合物在不同干燥时间内均能被检测到,表现相对稳定;(4) –5 ℃及17 ℃储存可保留更多挥发性化合物,室温保存则失去较多化合物,但主要的萜烯类化合物在不同储存方式下均可被检测到;(5) 90 ℃高温萃取可获得更多的挥发性化合物,但70 ℃萃取得到的萜烯类化合物相对含量较高,室温萃取所得化合物种类虽最少,但更能反映园林应用中枫香叶芳香物质挥发的实际情况。研究结果可为枫香叶的采后处理及高值化资源利用提供参考。     

联用SPME与GC-MS技术分析新鲜、萎蔫、干枯枫杨挥发性成分

运用SPME与GC/MS联用技术从新鲜枫杨叶挥发性物质中共检测出化合物52种,主要成分是橙花叔醇(15.54%)、吉马烯A(14.22%)、反式子丁香烯(10.19%)、7,8,9,10-四氢化-S-三氮唑(3,4-A)-呔嗪(8.59%)、十六酸(6.22%);萎蔫枫杨叶挥发性物质中共检测出化合物32种,主要成分是α-红没药烯(17.69%)、橙花叔醇(16.64%)、反式子丁香烯(14.26%)、α-桉叶烯(9.14%);干枯枫杨叶挥发性物质中共检测出化合物31种,主要成分是反式子丁香烯(23.30%)、α-红没药烯(17.75%)、α-桉叶烯(12.69%)、橙花叔醇(8.35%)、杜松烯(7.O%).随着枫杨萎蔫程度的变化,三者含有的化合物组分以及其相对含量都发生变化.本研究是首次报道枫杨的挥发性化学成分分析.     

施肥与摘蕾对棉蚜、棉叶蝉种群数量及棉花产量的影响

分析比较了不同施氮水平(0-450kg·hm^-2)与不同摘蕾处理(0-12个·株^-1)对转Bt基因棉花(GK-12)和常规棉花(泗棉3号)的棉蚜、棉叶蝉种群数量以及棉花产量的影响．结果表明,两个品种的棉蚜、棉叶蝉种群数量和有效铃量没有显著性差异。在本实验水平下,施肥和摘蕾与施肥的交互作用对两种棉花的棉蚜数量有显著性的影响,而摘蕾、施肥和两者的交互作用对两种棉花的棉叶蝉数量没有显著影响．在花铃期,施氮肥能增加两种棉花的有效铃数．线性回归分析表明,两种棉花的棉蚜、棉叶蝉的数量与施氮肥量和摘蕾量成正相关关系;有效铃量与施氮肥量呈正相关,与摘蕾量呈负相关．     

睡莲属62个栽培种花朵挥发性成分GC-MS分析北大核心CSCD

为了解睡莲花朵的致香物质,利用气相色谱-质谱法对62个栽培种花朵的挥发性成分进行了研究。结果表明,共检测出72种挥发性成分,以烯烃类(26种)、烷烃类(11种)和醇类(9种)较多,其中花香成分有53种(73.60%)。40个热带睡莲花朵中共检测出56种挥发性成分,其中花香成分39种;22个耐寒睡莲品种花朵共检测出37种挥发性成分,其中花香成分27种。花香成分中主要致香物质有乙酸苄酯、顺式-罗勒烯、苯甲醇、金合欢烯、月桂烯、柠檬烯、苯甲醛、α-异松油稀、α-蒎烯、肉桂醇和β-丁香醇等。利用组内联接余弦的方法,分别根据挥发性成分和花香成分,62个睡莲栽培种(品种)可分成3和4组。这为睡莲香气物质的开发利用及与传粉动物的协同进化研究提供了基础资料。     

盐胁迫对Bt棉棉蕾杀虫蛋白表达的影响

为探讨土壤盐分对转Bt基因抗虫棉棉蕾抗虫性的影响程度,采用盆栽试验,以2个Bt棉品种‘新棉33B’(盐敏感)和‘中07’(耐盐)为试验材料,设置5个土壤盐分水平(0、0.15%、0.30%、0.45%和0.60%), 研究盐分对蕾期Bt棉棉蕾中杀虫蛋白含量、Bt基因相对表达量及氮代谢相关酶和物质代谢的影响.结果表明: 棉蕾中Bt杀虫蛋白含量随土壤盐分的升高而降低,与对照相比(0%),2个不同类型棉花品种在土壤盐分0.30%及以上时,棉蕾中杀虫蛋白含量显著下降,且随土壤盐分水平的升高,Bt 杀虫蛋白含量下降幅度增大.土壤盐分胁迫提高了棉蕾中Bt基因的相对表达量.在相同土壤盐分水平下,盐敏感品种棉蕾中Bt杀虫蛋白含量受土壤盐分影响较大.杀虫蛋白表达量下降幅度大的品种,其可溶性蛋白含量、谷氨酸丙酮酸转氨酶(GPT)和谷氨酸草酰乙酸转氨酶(GOT)活性下降幅度较大,游离氨基酸含量、蛋白酶和肽酶活性上升幅度也较高.说明土壤盐分胁迫下,Bt棉棉蕾蛋白质合成能力下降,分解能力增强,可能是导致杀虫蛋白表达量下降的主要原因.     

田间条件下不同诱导棉花挥发性物质的鉴定

采用人工机械损伤(处理A)、棉铃虫为害(处理B)、棉铃虫为害+棉铃虫体(处理C)和水杨酸诱导(处理D)4种处理方式,对田间现蕾期棉花植株挥发性化合物进行了定性和定量研究.结果表明：有30多种棉株挥发性化合物被定性检出,对其中10种主要挥发性物质进行定量分析,田间棉株挥发的绿叶气味化合物有3-己酮、2-己酮和3-己醇等;单萜类化合物有α-蒎烯、β-蒎烯、β-月桂烯、3,7-二甲基,1,3,6-辛三烯等;脂肪族化合物有丙酸丁酯、乙酸戊酯、乙酸丁酯和3-甲基丁酸乙酯;芳香族化合物有苯乙酮、苯甲醛等.处理C和处理B的棉花挥发信息化合物种类和释放量均明显多于对照,这两个处理检测到3,7-二甲基,1,3,6-辛三烯;而处理A和处理D未检测到该化合物.与对照相比,处理A挥发物在种类和含量上差异不显著.在田间,处理B、处理C和处理D均能诱导促进棉株挥发物的释放.     

外源基因导入对棉花烟粉虱种群的影响

以转基因棉花和对应的常规棉花亲本为材料,探讨外源基因导入对棉花烟粉虱Bemisia tabaci(Gennadius)种群的影响。结果表明,在转基因棉花GK12、33B、SGK上,烟粉虱从卵发育到成虫羽化的历期分别为15.56、15.35和15.25 d,较对应的常规棉亲本SM3(19.38 d)、33(20.81 d)、SY321(18.76 d)分别短24.55%、26.23%、18.71%;GK12和33B棉花上烟粉虱的存活率(69.16%)分别较对应的常规棉亲本SM3(54.76%)和33(64.91%)高26.29%和12.81%,而SGK(63.21%)上烟粉虱的存活率与对应的常规棉亲本SY321(62.61%)之间差异不显著;烟粉虱在GK12(84.00)和33B(77.25)上的产卵量分别较SM3(62.25)、33(70.00)高34.93%和10.35%,但SGK和SY321之间差异不显著;在转基因棉花GK12、33B、SGK上,烟粉虱的雌雄性比分别比对应的常规棉亲本高26.71%、46.23%和19.17%。结果表明,外源基因导入后,有助于烟粉虱的发育,提高了烟粉虱的产卵量和雌雄性比,从而促进了烟粉虱种群的上升。     

转基因棉Bt毒蛋白含量与抗棉大卷叶螟效果间的关系分析

棉大卷叶螟Sylepta derogata Fabricius为近年来长江流域棉花中后期的一种重要害虫。在其危害盛期测定了8种转基因棉叶片Bt毒蛋白的含量与受害程度,在此基础上就转基因棉对棉大卷叶螟的抗虫性采用不同抗性指标相结合的方法进行了综合评估,同时对转基因棉对棉大卷叶螟的抗虫效果与毒蛋白含量的相关性进行分析。结果表明:各转基因棉品种的不同部位叶片的毒蛋白含量呈现顶叶最高,功能叶次之,老叶含量最低的趋势。抗蚜8017和SGK321 2个转基因棉品种的棉叶毒蛋白表达量相对较低,平均值在120μg/g以下,对棉大卷叶螟的抗性级别均为中抗。其它供试转基因棉棉叶毒蛋白表达量均值在150μg/g以上,抗性级别也均为高抗,而非转基因棉泗棉3号与石远321对棉大卷叶螟不具抗性。转基因棉对棉大卷叶螟的抗性水平与毒蛋白含量呈正相关。     

转Bt基因抗虫棉对棉蚜的杀虫剂敏感性及解毒酶系的影响

采用浸叶法,以亲本常规棉作对照,室内测定了吡虫啉和辛硫磷对在转Bt基因棉上取食不同世代棉蚜Aphis gossypii Glover的毒力,并用酶标仪测定了转Bt基因棉对棉蚜酯酶、羧酸酯酶和乙酰胆碱酯酶活力的影响。结果表明,棉蚜对吡虫啉和辛硫磷的敏感性并未受Bt棉的影响,2种杀虫剂对分别取食Bt棉1代和多代（60代以上）的棉蚜与取食亲本常规棉棉蚜的LC₅₀值之间均无显著差异。在抗虫棉GK12上取食1代和21代的棉蚜,其酯酶、羧酸酯酶和乙酰胆碱酯酶的活力与取食亲本常规棉的棉蚜之间差异不显著。可见,转Bt基因棉对棉蚜的杀虫剂敏感性和解毒酶活力无显著影响。     

Effects of transgenic Bt cotton on the population density, oviposition behavior, development, and reproduction of a nontarget pest, Adelphocoris suturalis (Hemiptera: Miridae)

Transgenic cotton has shown great promise for the control of target pest insects; however, frequent outbreaks of nontarget pest mirids has been recorded in recent years in northern China. To test the hypothesis that transgenic cotton contributes to nontarget pest outbreaks, we studied the impact of transgenic Bt cottons (both Bt and Bt + CpTI) on the fitness of nontarget pest Adelphocoris suturalis Jakovlev. No significant differences were detected between population densities of A. suturalis in unsprayed nontransgenic cottons and in unsprayed transgenic Bt cottons in 2007, 2008, and 2009. No difference in preferred oviposition site or egg production was detected between transgenic and nontransgenic cottons in both free choice and no choice tests. No difference in life table parameters was detected for A. suturalis between Bt cottons and nontransgenic cottons. All these results indicated that transgenic crops did not contribute to the nontarget pest outbreaks when being compared with their parental lines. The possible reasons for intensified pest status of A. suturalis, such as decrease of pesticide application, deficient natural enemies, and area-wide shift of cotton varieties, were discussed.     

Effect of pyramiding Bt and CpTI genes on resistance of cotton to Helicoverpa armigera (Lepidoptera: Noctuidae) under laboratory and field conditions

Transgenic cotton (Cossypium hirsutum L.) varieties, adapted to China, have been bred that express two genes for resistance to insects, the CrylAc gene from Bacillus thuringiensis (Berliner) (Bt), and a trypsin inhibitor gene from cowpea (CpTI). Effectiveness of the double gene modification in conferring resistance to cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), was studied in laboratory and field experiments. In each experiment, performance of Bt+CpTI cotton was compared with Bt cotton and to a conventional nontransgenic variety. Larval survival was lower on both types of transgenic variety, compared with the conventional cotton. Survival of first-, second-, and third-stage larvae was lower on Bt+CpTI cotton than on Bt cotton. Plant structures differed in level of resistance, and these differences were similar on Bt and Bt + CpTI cotton. Likewise, seasonal trends in level of resistance in different plant structures were similar in Bt and Bt+CpTI cotton. Both types of transgenic cotton interfered with development of sixth-stage larvae to adults, and no offspring was produced by H. armigera that fed on Bt or Bt+CpTI cotton from the sixth stage onward. First-, second-, and third-stage larvae spent significantly less time feeding on transgenic cotton than on conventional cotton, and the reduction in feeding time was significantly greater on Bt+CpTI cotton than on Bt cotton. Food conversion efficiency was lower on transgenic varieties than on conventional cotton, but there was no significant difference between Bt and Bt+CpTI cotton. In 3-yr field experimentation, bollworm densities were greatly suppressed on transgenic as compared with conventional cotton, but no significant differences between Bt and Bt+CpTI cotton were found. Overall, the results from laboratory work indicate that introduction of the CpTI gene in Bt cotton raises some components of resistance in cotton against H. armigera, but enhanced control of H. armigera under field conditions, due to expression of the CpTI gene, was not demonstrated.     

不同类型品种棉花上棉蚜适生性及种群动态

通过田间调查、室内网罩盆栽苗测定选择性等方法,考察了常规棉（泗棉3号,石远321）、杂交抗虫棉（辽棉19号,鲁棉研18号）、转单价基因抗虫棉（国抗12号,中棉所32）和转双价抗虫棉（SGK321,中棉所41）4种类型8个品种棉花上棉蚜的适生性及种群动态。结果表明： 棉蚜在各棉花品种上的种群动态有明显差异（P<0.05）,单株蚜量以转单价基因抗虫棉中棉32上最高,常规棉泗棉3号上最低,分别为297.81头/株和76.88头/株。棉蚜对4种类型棉花品种的选择性有明显差异（P<0.05）,其中对转单价基因抗虫棉有很强的选择性。根据棉蚜实验种群的参数判断,其在不同品种棉花上的生长发育、存活及繁殖存在显著差异： 若虫发育历期常规棉石远321上最长（6.46天）,双价棉中棉所41上最短（5.75天）; 存活率转单价基因抗虫棉中棉32上最高（88.21%）,双价棉SGK321上最低（76.46%）; 单雌产蚜量杂交抗虫棉辽棉19上最大（44.48头）,双价棉SGK321上最小（33.51头）; 内禀增长率转单价基因抗虫棉中棉32上最高（0.3695）,双价棉中棉所41上最低（0.3389）。综合评价,棉蚜的生存和繁殖适合性在转单价基因抗虫棉上最高,在双价棉上最低。     

Impact of transgenic Bacillus thuringiensis cotton on a non-target pest Tetranychus spp. in northern China

Field studies were carried out to evaluate the effect of two transgenic cotton varieties (SGK321 carrying Cry1A+CpTI and DP99B carrying Cry1Ac) and the conventional variety (shiyuan321‐parental line of SGK321) on spider mites, Tetranychus spp. from 2002 to 2004. In 2002, this experiment included three treatments: Bt cotton field (SGK321) treated with acaricides against spider mites, untreated non‐Bt cotton field (Shiyuan321), and untreated Bt cotton field (SGK321). In 2003–2004, there are four types of treatments after a new transgenic Bt cotton variety, DP99B (non‐chemical control), was added into the experiments. The results showed that there were no significant difference in densities of spider mites among Bt without acaricides and non‐Bt without acaricides cotton fields, nor was there a significant difference in damage of spider mites to cotton among these treatments (P > 0.1). However, there are significant differences (P < 0.05) in densities of spider mites and damage caused by spider mites between cotton fields with and without acaricides. Acaricide significantly reduced the densities of spider mites in Bt cotton (P < 0.05). These results suggest that Bt cotton has no effect on spider mites populations. However, spider mites have the potential for severe damage in Bt cotton fields. Acaricides are essential tools in controlling cotton spider mites in northern China.     

取食转Bt+CpTI基因棉花的棉蚜对普通草蛉生长发育及繁殖力的影响

【目的】研究取食转Bt+CpTI基因棉花的棉蚜Aphis gossypii Glover对普通草蛉Chrysopa carnea Stephens生长发育及繁殖力的影响,为转Bt基因抗虫棉花的推广和应用提供理论支持。【方法】普通草蛉取食转Bt+CpTI棉花SKG321和对照品种石远321棉花上的棉蚜后,对普通草蛉生长发育情况进行研究,并对其幼虫、蛹的存活率和成虫的羽化率及产卵量进行分析。【结果】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的各龄幼虫发育历期、蛹期和产卵前期无不利影响;处理与对照间各龄幼虫存活率、蛹的存活率和成虫羽化率之间没有显著差异;取食转Bt+CpTI棉花上棉蚜的普通草蛉单雌日产卵量与对照相比没有减少,而25 d内的单雌总产卵量与对照相比差异亦不显著。【结论】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的生长发育及繁殖力无负面影响。     

Antennal response of cotton bollworm (Heliocoverpa armigera) to volatiles in transgenic Bt cotton

Abstract:  Volatile compositions in transgenic Bt cotton variety 'GK-97' and its parental variety 'Simian No. 3' were analysed, and antennal response of cotton bollworm ( Heliocoverpa armigera ) to the volatiles were investigated. Volatile components in Bt cotton and regular cotton were almost same, except for absence of a sesquiterpene and a minor unknown compound in regular cotton. The results of gas chromatography-electroantennographic detection (GC-EAD) experiments showed that cotton bollworm antennae responded to seven compounds and two minor unknown compounds in Bt volatiles, among which concentrations of α -pinene and β -pinene in Bt cotton were relatively higher than those in regular cotton, and one minor unknown compound was absent in regular cotton.     

转基因抗虫棉Bt基因不同剂量的聚合与抗虫性表现

通过有性杂交手段培育出聚有不同数目Bt基因的植株,在不同生育期进行抗虫性测定和Bt毒蛋白表达的ELISA检测,旨在揭示聚合不同数目Bt基因的植株抗虫性的互作表达机理。聚合有1－4个Bt基因的植株在整个生育期的抗虫性、毒蛋白表达特性和单价抗虫棉时空表达一致,生育前期抗虫性好、毒蛋白表达量高;生育中、后期抗虫性有所下降,毒蛋白表达量降低。聚合有4个Bt基因的纯合材料并未因Bt基因的增加而起到抗性增强的效果,相反还因同源抑制而有所降低。不同来源的Bt基因处于杂合状态时其抗虫性和Bt毒蛋白量均得到充分表达。