5种杀虫剂对黑粪蚊的毒力及对平菇菌丝生长的影响

[目的]黑粪蚊为食用菌生产中的危险害虫之一,通过杀虫剂的毒力测定和对平菇菌丝生长抑制试验,拟筛选获得理想的黑粪蚊防治杀虫剂。[方法]采用毒饵法测定了5种杀虫剂对黑粪蚊幼虫的室内毒力,并研究各杀虫剂对平菇菌菌丝生长的抑制情况。[结果]20%甲氰菊酯乳油、14%阿维·虫螨腈悬浮剂和20%甲维·吡丙醚悬浮剂对黑粪蚊的校正死亡率达80%以上,对黑粪蚊幼虫均有较好的杀灭效果,而苏云金杆菌（以色列亚种）悬浮剂对黑粪蚊幼虫的杀虫效果不理想。5种杀虫剂对平菇菌丝生长均有一定抑制作用,且农药品种间存在极显著差异,20%甲氰菊酯乳油抑制率最高,高浓度480 mg·L^-1处理的抑制率可达16.85%,14%阿维·虫螨腈悬浮剂的抑制率最低,高浓度140 mg·L^-1处理的抑制率仅为5.83%。[结论]本研究中14%阿维·虫螨腈悬浮剂用于平菇菌中黑粪蚊防治效果最好,研究结果将为食用菌黑粪蚊的药剂防治提供理论指导。     

驱避色防治平菇蚊、蝇类害虫的效果评价

为探讨运用驱避色防治平菇Pleurotus ostreatus蚊、蝇类害虫的可行性,本研究基于一种对平菇双翅目害虫具明显驱避作用的浅灰色（RGB=169, 169, 169）用于设计菌袋、地膜和棚膜,以评估这种驱避色赋能平菇配套栽培设施（菌袋、地膜和棚膜）对平菇蚊、蝇类害虫的驱避效果和增产作用。采用正交设计各影响因子处理组合进行田间小区试验,研究不同菌袋、地膜、棚膜颜色对害虫驱避率及平菇增产率的影响,筛选出最优组合,应用于生产实际。结果表明,驱避色菌袋+驱避色地膜+透明棚膜的处理组合的害虫驱避率和增产率均最高,与对照组相比,驱避率增加了45.63%,第1、2、3潮菇产量和总产量分别增加了11.29%,2.55%,8.82%和7.74%。通过多因素和差异显著性分析,菌袋颜色是影响害虫驱避率和增产率的主因子,与对照相比对害虫驱避率和前3潮菇增产率差异达到极显著水平。在平菇生产过程中,使用驱避色菌袋和地膜能有效控制平菇蚊、蝇类害虫的危害并提高产量,是一项值得推广的害虫防治措施。     

不同生育期转Bt基因棉种植对根际土壤微生物的影响

 在盆栽种植条件下,比较研究了两个转Bt基因棉(Gossypium hirsutum)与对照棉对根际土壤细菌、放线菌、真菌和主要功能类群及多样性的影 响差异。结果表明：两个转Bt基因棉根际土壤均可检测到Bt蛋白,且不同转Bt基因棉根系分泌Bt蛋白量以及Bt蛋白在根际土壤中的降解率不同 。与各自对照相比,转Bt基因棉对细菌和真菌生长繁殖有促进作用,对放线菌、好气固氮菌和钾细菌数量没有显著影响。苗期和花期转Bt基因 棉均可显著提高氨化细菌、显著降低无机溶磷菌数量,花期均可显著提高好气纤维分解菌、显著降低有机溶磷菌数量,‘Bt冀668’苗期也可显 著提高好气纤维分解菌数量。转Bt基因棉根际土壤好气纤维分解菌、有机和无机溶磷菌多度发生了变化。尽管功能类群总数转Bt基因棉高于各 自对照常规棉,但群落多样性和均匀度都有所下降,优势集中性表现明显,且花期转Bt基因棉多样性参数值以及功能类群数量的变化幅度大于 苗期。     

Bt棉田边缘杂草带对棉田内叶螨发生的影响

2003～2004年,在冀南棉区系统研究了棉田边缘杂草带对棉田内叶螨发生的影响。试验设3个处理：转Bt基因棉化防田(使用杀螨剂控制棉叶螨且保留棉田边缘杂草)、转Bt基因棉对照田(保留棉田边缘杂草)和转Bt基因棉除草田(去除棉田边缘杂草)。结果表明,2003年6月28日～8月7日,处理间的叶螨发生量和有螨株率差异明显。2003年叶螨发生高峰期(7月28日),对照田的百株平均螨量达834头,分别是化防田、除草田的9.4倍和11.5倍;对照田的有螨株率的峰值为34.7%,分别高于化防田和除草田6.4%和12%;棉叶螨的总计值,对照田分别是化防田和除草田的5.8倍和5.5倍。2004年7月10日～8月29日,对照田的百株螨量和有螨株率明显高于除草田和化防田。2004年叶螨发生高峰期（8月10日）,对照田的百株平均螨量达1 222头,分别是化防田、除草田的4.3倍和23.4倍;对照田的有螨株率的峰值达100%,分别比除草田和化防田多75%和87%;棉叶螨的总计值,对照田分别是化防田和除草田的4.9和9.7倍。两年中,除草田和化防田的百株螨量和有螨株率峰值出现日期有所不同。对照田内,棉田边缘杂草至所调查的棉株距离同螨害指数呈显著(P＜0.05)或极显著(P＜0.01)的负相关。本研究表明,去除棉田边缘杂草的棉田,叶螨发生始期较晚且发生量较少;棉田边缘杂草到取样点的距离与棉叶螨的为害程度呈直线负相关,距离越近,叶螨的发生为害越重。     

转基因棉种植对土壤水解酶活性的影响

转Bt基因棉和转Bt＋cpTI基因棉种植面积不断扩大,它们种植后杀虫晶体蛋白在土壤中的残留特性及对土壤水解酶活性的影响是环境风险评价的重要组成部分,本文采用盆栽实验的方法对此进行了初步研究。结果表明,转基因棉出苗后生长到30天时可向土壤中释放Bt杀虫晶体蛋白,而双抗棉种植时CpTI杀虫晶体蛋白的释放量与品种有关;转基因棉出苗后30天时,与等价基因系非转基因棉（各对照）相比,转Bt基因棉（“中30”）和双抗棉A（转Bt＋CpTI棉“中41”）的种植并未使脲酶、蛋白酶和磷酸单脂酶活性发生显著变化,而双抗棉B（转Bt＋CpTI棉“双抗321”）的种植使土壤磷酸单脂酶活性显著下降。从杀虫晶体蛋白的释放和对酶活性的影响来看,双抗棉A的种植对土壤的生物活性扰动更小。     

转Bt基因棉对非靶标害虫及害虫天敌种群动态的影响

2000～2001年,在湖北棉区系统研究了转Bt基因棉对棉田非靶标害虫及天敌种群动态的影响。试验设三个处理：转Bt化防田（使用化学农药控制害虫）、转Bt自控田（仅依靠田间自然天敌控制害虫）及常规对照棉田（利用综合防治措施控制害虫）。结果表明,在转Bt基因棉田中,除棉蓟马外,其它主要非靶标害虫（主要是刺吸性害虫）的种群发生数量呈明显的上升趋势。2000年棉蚜发生的总计值,化防田和自控田分别比常规对照田增加37.9%和71.4%,2001年则分别增加92.5%和134.9%;2000年朱砂叶螨发生的总计值,化防田和自控田分别比常规对照田增加181.1%和298.3%,2001年则分别增加69.9%和105.0%;转Bt基因棉对于斜纹夜蛾与烟粉虱这两种近几年来对棉花危害有加重趋势的害虫,在大田中没有表现出抗性。天敌的发生种类和数量也都远远高于常规对照棉田,几种主要天敌种群发生的总计值及高峰值都明显高于常规对照棉田。2000年蜘蛛类发生的总计值,化防田和自控田分别比常规对照田增加66.3%和112.1%,2001年则分别增加95.1%和111.7%;2000年龟纹瓢虫发生的总计值,化防田和自控田分别比常规对照田增加140.8%和135.4%,2001年则分别增加67.2%和109.5%;2000年在两块转Bt基因棉田中种群数量较大的大眼蝉长蝽在常规对照棉田的调查过程中没有发现,2001年在对照田中也仅记录到1头。试验表明,充分利用Bt棉田中天敌种类较丰富、种群数量较大的优势而加强生物防治,是湖北棉区转Bt基因棉田综防体系中的一个关键措施。     

应用电击法获得转MT基因平菇

将MT基因用电击法转化平菇(Pleurotus ostreatus),MT基因表达蛋白与金属离子结合而形成络合物,用Zn 诱导,转基因平菇能富集Zn,可为缺Zn的人群补充Zn,使平菇成为一种保健和治疗的食品或蔬菜.原生质体制备浓度为6.745×106个/mL.原生质体电击转化率为0.01%.PCR检测,200 bp处有MT基因条带.蛋白检测:转基因MT平菇ELISA检测阳性,表达率为0.6%～0.8%.SDS-PAGE 显示有表达条带.Western blot显示有阳性条带.抗ZnSO4结果:野生型平菇抗ZnSO4浓度为1.0 mmol/L,1.2 mmol/L开始受抑制,转基因平菇抗ZnSO4浓度为1.5 mmol/L, 2.0 mmol/L开始受抑制. 出菇试验结果表明,在米糠与锯沫比为1∶3的培养基上生长,在米糠与锯沫比为1∶4的培养基上不生长.24 d菌丝可在广口瓶中长满,用于子实体培养.     

湖北棉区转Bt基因棉对棉铃虫的控制作用

2000-2001年通过田间系统调查,表明转Bt基因棉（品种为GK19）在湖北江汉平原棉区对棉铃虫抗性稳定。试验设3个处理：转Bt基因棉化防田（使用化学农药控制害虫）、转Bt基因棉自控田（依靠天敌控制害虫）及常规棉对照田（利用综合防治措施控制害虫）。从棉铃虫的第2代到第5代整个发生期内,即使在不进行化学防治的情况下,棉铃虫在Bt棉田的发生量也保持在极低的水平(最高百株虫量为12头)。室内饲养结果表明,转Bt基因棉对棉铃虫的生长发育（幼虫体重、蛹重）有较为明显的影响,使6龄幼虫体重减少25.6%,蛹重减少18.2%。棉铃虫幼虫取食转Bt基因棉组织后,发育迟缓,相对于常规棉喂养的整个发育历期延长17 d,使棉铃虫在田间的危害减少至少一个世代。另外,接虫试验表明,棉铃虫幼虫在常规棉上的取食时间是转Bt基因棉株上的6.1倍,极大地减轻了棉铃虫的危害程度。     

Bt抗虫棉秸秆还田对土壤养分特征的影响

【目的】研究转基因作物秸秆或残茬还田可能对土壤养分特性造成的影响。【方法】以不同抗虫水平Bt棉花和常规棉花(泗棉3号)为研究材料,分别在经过一、二个生长周期后将秸秆机械粉碎后原位还田,40 d后测定分析土壤中Bt蛋白含量及肥力相关的养分含量变化。【结果】Bt棉秸秆还田后,所有品种棉花土壤中Bt蛋白含量与还田前无显著增加,且转Bt基因棉与非转基因棉还田对土壤Bt蛋白含量的影响并无显著差异。同时,棉秸秆还田可显著提高土壤有机质、速效磷、碱解氮、速效钾、全氮、全磷和全钾含量,提升土壤pH值;增加幅度在不同抗虫水平Bt棉花间及与非转基因常规棉花品种间皆无显著性差异。【结论】秸秆还田对土壤肥力的提升与Bt棉的抗虫水平无关。“转Bt基因”不成为Bt棉秸秆还田提高土壤肥力的限制性因素,其秸秆还田不会对土壤肥力质量产生负面影响,可使土壤养分含量增加,有效提升土壤肥力。秸秆原位还田简单、无害又提升肥力,有条件作为转Bt基因植物秸秆无害化处理的理想方式。     

转CpTI-Bt基因棉和转Bt基因棉对棉铃虫幼虫存活、生长及营养利用的影响

2000年7月中旬和8月中旬, 分别测定了采自田间的转CpTI-Bt基因双价抗虫棉（SGK321, 以下简称CpTI-Bt棉）和转Bt基因抗虫棉（中30,以下简称Bt棉）对棉铃虫Helicoverpa armigera幼虫存活、生长的影响。结果表明：7月中旬两种转基因抗虫棉抗虫效果均较好,尤其是CpTI-Bt棉棉叶和花瓣对4龄幼虫3天内致死率为92%以上;8月中旬两种转基因棉的抗虫活性均明显降低,且Bt棉的杀虫活性显著低于CpTI-Bt棉,其幼虫死亡率与对照受体棉中16的死亡率之间无显著差异,仅显著抑制了幼虫的生长;石远321(SGK321受体品系)的花瓣具有一定的抗虫活性,可显著降低取食幼虫的体重,甚至造成部分幼虫死亡; CpTI-Bt棉中,花瓣和棉叶的抗虫性明显高于蕾和铃心。对5龄幼虫取食棉铃1日后的营养指标测定结果显示： 两种转基因抗虫棉处理的幼虫相对生长率和相对取食量均显著低于石远321,但两者之间无显著差异; CpTI-Bt棉处理的幼虫近似消化率显著低于石远321和Bt棉,但其食物利用率显著高于石远321和Bt棉。     

The substantive equivalence of transgenic (Bt and Chi) and non-transgenic cotton based on metabolite profiles

Compositional studies comparing transgenic with non-transgenic counterpart plants are almost universally required by governmental regulatory bodies. In the present study, two T₂ transgenic cotton lines containing chitinase (Line 11/57) and Bt lines (Line 61) were compared with non-transgenic counterpart. To do this, biochemical characteristics of leaves and seeds, including amino acids, fatty acids, carbohydrates, anions, and cations contents of the studied lines were analyzed using GC/MS, high-performance liquid chromatography (HPLC), and ion chromatography (IC) analyzers, respectively. polymerase chain reaction (PCR) and Western blot analyses confirmed the presence and expression of Chi and Bt genes in the studied transgenic lines. Although, compositional analysis of leaves contents confirmed no significant differences between transgenic and non-transgenic counterpart lines, but it was shown that glucose content of chitinase lines, fructose content of transgenic lines (Bt and chitinase) and asparagine and glutamine of chitinase lines were significantly higher than the non-transgenic counterpart plants. Both the transgenic lines (Bt and chitinase) showed significant decrease in the amounts of sodium in comparison to the non-transgenic counterpart plants. The experiments on the seeds showed that histidine, isoleucine, leucine, and phenylalanine contents of all transgenic and non-transgenic lines were the same, whereas other amino acids were significantly increased in the transgenic lines. Surprisingly, it was observed that the concentrations of stearic acid, myristic acid, oleic acid, and linoleic acid in the chitinase line were significantly different than those of non-transgenic counterpart plants, but these components were the same in both Bt line and its non-transgenic counterpart. It seems that more changes observed in the seed contents than leaves is via this point that seeds are known as metabolites storage organs, so they show greater changes in the metabolites contents comparing to the leaves.     

NaCI胁迫对转基因杨树外源基因表达的影响

以具高抗虫性的转抗虫基因‘74l杨’及在此基础上转入了发根农杆菌融质粒T-DNA株系的组培苗为材料,研究了转基因株系BtCrylAc抗虫基因和发根基因的表达及其对NaCI胁迫的反应。结果表明,转入Ri质粒T-DNA上的rol基因后,导致苗木根系数目增加,根系长度减小,IAA和GA含量显著提高,抗虫BtCrylAc基因编码的毒蛋白的表达量降低;随着NaCI胁迫强度的增加,苗高、根系数量、叶绿素含量及IAA、GA含量逐步降低,而根系的长度加大,Bt毒蛋白含量显著提高,表明NaCI胁迫使转基因杨外源Bt毒蛋白基因的表达增强,而发根农杆菌．Ri质粒T-DNA的表达下降。     

大田环境下转Bt基因玉米对土壤酶活性的影响

在大田自然条件下,比较研究了转Bt基因玉米和非转基因亲本玉米在种植和秸秆分解时对土壤酶活性影响的差异。结果表明,与亲本非转基因玉米相比,在各生育期内种植转Bt玉米对土壤蛋白酶和土壤脲酶活性均没有显著影响;在喇叭口期和抽雄期,土壤蔗糖酶和土壤酸性磷酸酶活性显著提高。在秸秆还田后,两种玉米秸秆对土壤酸性磷酸酶活性的影响没有显著差异,但使用转Bt玉米秸秆的土壤蔗糖酶、土壤脲酶和土壤蛋白酶的活性则有显著提高。与亲本玉米相比,在所有观测期内,种植Bt玉米及秸秆还田对土壤酶活性的影响,在影响的幅度及趋势上随玉米生育期和土壤酶种类的不同而产生差异,但没有观测到显著不利影响;商业化Bt玉米的环境释放仍有待长期定位观测和评价。     

转Bt基因水稻根际土壤微生物多样性的磷脂脂肪酸(PLFAs)表征

以亲本水稻为对照,应用13C脉冲标记和磷脂脂肪酸技术,分析转Bt基因对水稻根际微生物多样性的影响.结果表明:转Bt基因水稻与亲本水稻根际均以饱和脂肪酸和支链脂肪酸为主,单不饱和脂肪酸次之,多不饱和脂肪酸最少.苗期、拔节期和抽穗期,转成基因水稻根际革兰氏阳性菌(G+)代表性磷脂脂肪酸含量显著低于亲本水稻;革兰氏阴性菌(G-)代表性磷脂脂肪酸含量显著高于亲本水稻.水稻各生育期,转Bt基因未对水稻根际土壤真菌、放线菌磷脂脂肪酸含量造成显著影响,且转Bt基因水稻与亲本水稻根际微生物磷脂脂肪酸13C含量无显著性差异.表明外源Bt基因插入仅对水稻根际微生物多样性造成短暂影响,不具有持续性.     

转基因棉花对棉铃虫天敌寄生率的影响

采用分期接虫、分期回收法研究常规棉与转基因棉花品种 (系 )上棉铃虫寄生性天敌在不同棉花品种上的寄生作用。结果表明 ,无论是第 2代棉铃虫发生期间还是第 3 ,4代棉铃虫发生期间 ,无论是棉铃虫卵期还是幼虫期 ,转基因棉花品种 (系 )的棉铃虫卵、幼虫寄生率均显著低于常规棉花品种上的寄生率。棉铃虫卵期寄生蜂主要是拟澳洲赤眼蜂 ,幼虫寄生蜂为棉铃虫齿唇姬蜂等。由此看出转基因棉对棉铃虫寄生性天敌存在非亲和性。     

RNAi‐mediated Ultra‐low gossypol cottonseed trait: performance of transgenic lines under field conditions

Cottonseed remains a low‐value by‐product of lint production mainly due to the presence of toxic gossypol that makes it unfit for monogastrics. Ultra‐low gossypol cottonseed (ULGCS) lines were developed using RNAi knockdown of δ‐cadinene synthase gene(s) in Gossypium hirsutum. The purpose of the current study was to assess the stability and specificity of the ULGCS trait and evaluate the agronomic performance of the transgenic lines. Trials conducted over a period of 3 years show that the ULGCS trait was stable under field conditions and the foliage/floral organs of transgenic lines contained wild‐type levels of gossypol and related terpenoids. Although it was a relatively small‐scale study, we did not observe any negative effects on either the yield or quality of the fibre and seed in the transgenic lines compared with the nontransgenic parental plants. Compositional analysis was performed on the seeds obtained from plants grown in the field during 2009. As expected, the major difference between the ULGCS and wild‐type cottonseeds was in terms of their gossypol levels. With the exception of oil content, the composition of ULGCS was similar to that of nontransgenic cottonseeds. Interestingly, the ULGCS had significantly higher (4%–8%) oil content compared with the seeds from the nontransgenic parent. Field trial results confirmed the stability and specificity of the ULGCS trait suggesting that this RNAi‐based product has the potential to be commercially viable. Thus, it may be possible to enhance and expand the nutritional utility of the annual cottonseed output to fulfil the ever‐increasing needs of humanity.     

Mineral element distribution in organs of dual-toxin transgenic (Bt+CpTI) cotton seedling

Abstract

Being a new cultivar, the physiology of transgenic cotton, especially dual-toxin transgenic (Bt+CpTI) cotton, is not yet completely understood. Twelve elements in three organs of dual-toxin transgenic cotton seedlings were analyzed by ICP-MS. The distributions of the 12 elements were substantially different from those of non-transgenic cotton. In particular, the contents of B, Mg, P, K and Ca were the highest in leaves, while those of Si, Fe, Rb and Cu were the highest in roots; other elements had similar contents in the two organs, which were higher than those in the stem. Compared with non-transgenic cotton, the 12 elements could be classified into four groups according to their contents and distributions in the three organs: (a) P, K and Cu: their contents in transgenic cotton were remarkably lower, especially contents of P and K in leaves that were one times lower than those in leaves of non-transgenic cotton; (b) B, Mg and Mo: their contents in leaves and roots of transgenic cotton were higher, but lower in stems, compared with non-transgenic cotton; (c) Si, Mn, Fe, Rb and Zn: compared with non-transgenic cotton, these were lower in leaves and stems, but higher in roots of transgenic cotton; and (d) Ca: compared with non-transgenic cotton, its content was higher in all three organs of the transgenic counterpart. The decrease in soluble proteins and the expression of Bt and CpTI genes could be responsible for these changes. Further studies are needed to verify this hypothesis.     

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。     

Growth and nutritional trials on transgenic Nile tilapia containing an exogenous fish growth hormone gene

In a long-term growth trial, transgenic tilapia Oreochromis niloticus L. showed a 2·5-fold increase in growth compared with non-transgenic siblings. At 7 months, mean mass of transgenic tilapia was 653 g compared with 260 g for non-transgenic siblings. A significant increase ( P >0·01) in head: total length ratio, viscera-somatic index and hepato-somatic index was observed in transgenic fish. Female gonado-somatic index ( I G) was found to be lower in transgenics than non-transgenic siblings in both mixed and separate culture conditions. Transgenic male I G values were found to be higher in mixed culture and lower in separate culture than that of their non-transgenic siblings. Food conversion efficiency was more than 20% greater in the transgenic fish. In a second shorter-term growth performance trial, the transgenic fish grew to about four times the size of their non-transgenic siblings. A digestibility trial suggested that transgenic tilapia were more efficient utilizers of protein, dry matter and energy. Apparent digestibility of protein and apparent energy digestibility were higher in the transgenic fish.