转基因抗虫玉米对亚洲玉米螟的抗性评价

采用田间人工接虫鉴定方法,根据玉米抗螟性田间鉴定评价标准,研究转基因玉米Bt799和NC6304YGRR对亚洲玉米螟的抗性.心叶期接虫试验结果表明,以食叶级别作为评价参数,2种转基因玉米对亚洲玉米螟具有良好的抗性,食叶级别均为1级;2种非转基因玉米食叶级别均大于7级.吐丝期接虫试验结果表明,以单株虫孔数、单株活虫数、单株隧道个数及单株隧道长度作为评价参数,2种转基因玉米对亚洲玉米螟抗性均显著高于各自对应的非转基因玉米.转基因抗虫玉米Bt799和NC6304YGRR田间抗虫效果良好,均能保护玉米在整个生育期内不受亚洲玉米螟危害.     

转Bt毒蛋白基因玉米及其抗虫性研究进展

本文从转Bt毒蛋白基因玉米的培育及商品化，Bt毒蛋白基因在转基因玉米中的遗传分离与整合、对玉米螟及其它害虫的杀虫效果、对天敌种群数量和玉米病害发生程度的影响、玉米螟对转Bt毒蛋白基因玉米产生抗性及解决措施、应用转Bt毒蛋白基因玉米潜在的生态风险性等方面对国内外最新研究进展进行了综述。     

玉米转基因研究进展

玉米转基因的方法主要有农杆菌、基因枪、PEG介导等方法,受体主要是愈伤组织和幼胚等生活较旺盛的部位。目前在玉米上的成功应用是抗玉米螟的Bt转基因玉米、抗除草剂转基因玉米等种类。转基因技术目前面临的问题是关于环境和食物链的问题。也就未来玉米的转基因发展进行了讨论。     

转Bt基因玉米对亚洲玉米螟的抗性及其农艺性状研究

以普通玉米杂交组合N3030、N4640为对照,通过盆栽接虫试验和田间调查,系统研究了转Bt基因玉米杂交组合N3030Bt和N4640Bt对亚洲玉米螟的抗虫效果和主要农艺性状变化。结果表明,转Bt基因玉米对亚洲玉米螟产卵无驱避作用,但花叶率、茎秆和雌雄穗的受害程度均明显轻于对照;转Bt基因玉米的生育期、株高、穗位高、穗长、穗粗等与对照差异不大,秃尖长明显短于对照,穗粒数和产量明显高于对照。     

转Bt基因玉米对亚洲玉米螟的抗性及其农艺性状研究

以普通玉米杂交组合N3030、N4640为对照，通过盆栽接虫试验和田间调查，系统研究了转Bt基因玉米杂交组合N3030Bt和N4640Bt对亚洲玉米螟的抗虫效果和主要农艺性状变化。结果表明，转Bt基因玉米对亚洲玉米螟产卵无驱避作用，但花叶率、茎秆和雌雄穗的受害程度均明显轻于对照；转Bt基因玉米的生育期、株高、穗位高、穗长、穗粗等与对照差异不大，秃尖长明显短于对照，穗粒数和产量明显高于对照。     

分子改造Cry蛋白基因的转基因玉米创制及其抗虫功能评价

利用结构域交换和密码子优化方法对苏云金芽孢杆菌(Bt)杀虫晶体蛋白Cry1Ab进行合理化设计改造,获得新型Bt蛋白编码基因cry FLIa。利用农杆菌介导法将该基因转入玉米Hi II中,对转基因后代开展分子鉴定和抗性评价等相关工作。结果表明,cryFLIa基因已整合进玉米基因组中,连续3代自交材料检测显示,转基因玉米目标基因正常表达并稳定遗传;蛋白在植株叶片中发育各时期的表达各异,在灌浆期蛋白表达量较高(471.883 ng/g)。室内生测和田间人工接虫试验表明,转cryFLIa基因玉米具有抗亚洲玉米螟的能力。     

转Bt基因玉米的抗螟性及产量分析

在玉米心叶期和穗期对3个转Bt基因玉米品种及其相对应的受体品种进行人工接虫,分析了转Bt基因玉米对玉米螟的抗性及对产量损失的影响.结果表明:转Bt基因玉米在玉米心叶期,对一代玉米螟表现为高抗;在穗期,对二代玉米螟也表现高抗.在接虫条件下,转Bt基因玉米可以减少产量损失30%以上.因此.种植转Bt基因玉米具有明显增产作用和经济效益.     

转基因玉米Bt蛋白降解规律及其与土壤动物互作研究

以转Cry11Ac基因玉米及其非转基因对照为试验材料,利用大、中、小不同孔径的凋落物分解袋,研究黑土区转基因玉米外源基因表达Bt蛋白的降解规律及其与土壤动物的互作关系。结果表明,第二年春天大孔径凋落物分解袋内转基因玉米残体Bt蛋白含量为初始含量的25.98%,至8月份Bt蛋白只剩下初始含量的0.35%。分解袋孔径对转基因玉米及其对照玉米凋落物分解率和Bt蛋白降解率均有显著影响,表现为大孔径中孔径小孔径,说明大型和中型土壤动物都可以促进凋落物分解和Bt蛋白的降解。与对照玉米相比,转基因玉米凋落物分解率和凋落物内土壤动物群落结构参数都没有显著差异(P0.05,t-test),说明导入Bt基因对玉米残体分解速率和土壤动物均无显著影响。     

转Bf基因玉米的抗螟性及产量分析

在玉米心叶期和穗期对3个转Bt基因玉米品种及其相对应的受体品种进行人工接虫,分析了转Bt基因玉米对玉米螟的抗性及对产量损失的影响。结果表明：转Bt基因玉米在玉米心叶期,对一代玉米螟表现为高抗;在穗期,对二代玉米螟也表现高抗。在接虫条件下,转成基因玉米可以减少产量损失30％以上。因此,种植转Bt基因玉米具有明显增产作用和经济效益。     

转基因玉米SW12-859的抗螟性及农艺性状评价

SW12-859为吉林省农业科学院农业生物技术研究所培育的转基因玉米杂交组合,全株表达Cry1F基因杀虫蛋白。本研究以其为材料,与其遗传背景相似的郑单958为非转基因对照,在玉米心叶期(6~8叶期)和穗期(13~16叶期)进行田间人工接玉米螟初孵幼虫,调查其抗螟性、农艺性状及对产量损失的影响。结果表明,在玉米心叶期和穗期,转基因玉米SW12-859对一、二代玉米螟表现为高抗,郑单958表现为感,二者受玉米螟危害程度(抗螟性)差异显著。转基因玉米SW12-859和郑单958的株高、穗位高等农艺性状相近,在接虫条件下,转基因玉米产量显著高于非转基因玉米。     

转Bt基因玉米的抗螟性及产量分析

在玉米心叶期和穗期对3个转Bt基因玉米品种及其相对应的受体品种进行人工接虫,分析了转Bt基因玉米对玉米螟的抗性及对产量损失的影响。结果表明：转Bt基因玉米在玉米心叶期,对一代玉米螟表现为高抗;在穗期,对二代玉米螟也表现高抗。在接虫条件下,转Bt基因玉米可以减少产量损失30%以上。因此,种植转Bt基因玉米具有明显增产作用和经济效益。     

Economics of long-term IPM for western corn rootworm

We used a published biological model and published economic algorithm to evaluate western corn rootworm (Diabrotica virgifera virgifera) IPM for growers over a 15 year period beginning after significant adoption of insecticidal corn (Zea mays). The primary focus of our analysis was the economic evaluation (grower profit) of transgenic insecticidal corn expressing Cry34/35Ab1 as event 59122 and its refuge planted continuously year after year (continuous corn). We chose the reference scenario for economic comparison to be the use of soil insecticides on continuous, conventional corn. The model simulated the evolution of rootworm resistance to transgenic insecticidal (Bt) corn; but did not simulate resistance to soil insecticides. We evaluated refuge sizes of 5–50% for single-trait Bt corn and 5–20% for pyramided Bt corn with two traits targeting western corn rootworm. We considered the role of block and blended (seed mixture) refuges for insect resistance management (IRM). Results demonstrated that, for pyramided Bt corn, block refuges planted in the same location within a field year after year gave the greatest overall profit for grower. If growers relocated their block refuge annually (which is the most common practice), then a 5% blended refuge gave the greatest return. For single-trait Bt corn, 10–20% blended refuges gave greater economic return compared to block refuges ranging from 5% to 50%. Single-trait Bt corn with 5–20% block refuge (with no insecticide) was superior to soil insecticide use alone in all cornfields.     

Sequential sampling plan for assessing corn rootworm (Coleoptera: Chrysomelidae) larval injury to Bt maize

In-field product performance assessments are an essential component of corn rootworm (Diabrotica spp.; CRW) resistance management plans for transgenic maize (Zea mays L.) products expressing proteins derived from the bacterium Bacillus thuringiensis (Bt). The goal of a successful field sampling program is to accurately characterize in-field product performance while also minimizing resource demand, as collection of maize root samples to evaluate CRW injury can present resource challenges such as labor intensiveness, potential safety issues, and a limited time window available for sampling. A resource-efficient sequential sampling plan was developed that utilizes data-driven root injury threshold values derived from benchmark product performance data for both single and pyramided Bt maize traits for CRW control. This sequential sampling methodology incorporates unbiased sampling and controlled false positive and false negative error rates, enabling accurate assessment decisions to be made with efficient resource use. Our proposed approach enables systematic and effective classification of in-field Bt maize product performance, with applications to other CRW control technologies besides Bt maize products.     

白僵菌与苏云金芽胞杆菌水悬浮剂研制及田间防治玉米螟研究

通过室内生物测定,研究球孢白僵菌和苏云金芽胞杆菌联合防治玉米螟的效果。研究表明,确定白僵菌与苏云金芽胞杆菌浓度分别为1.0×106孢子/mL、1.0×109孢子/mL条件下,杀虫效果达到最佳,LT90为6.64(±0.07)d,较1.0×109的Bb LT90缩短26.06%。通过助剂悬浮率、乳化效果以及助剂对微生物活性影响等确定1个白僵菌与苏云金芽孢杆菌水悬浮剂剂型配方。田间防治玉米螟结果表明,两种生防菌均在防治过程中起作用,田间玉米受害株数、活虫数均明显低于未施药对照,防治效果优于单一使用白僵菌或苏云金芽胞杆菌。     

Comparative assessment of the field-susceptibility of Sesamia nonagrioides to the Cry1Ab toxin in areas with different adoption rates of Bt maize and in Bt-free areas

A three year (2003-2005) field study compared the susceptibility to the Cry1Ab toxin, expressed in Bt maize, of Mediterranean corn borer (MCB) Sesamia nonagrioides populations collected from areas with different adoption rates of Bt maize in Spain with Bt-free areas in Greece. Spain is the only European country where the cultivar Compa CB derived from the event Bt176 was commercially grown, from 1998 to 2005. The large decrease of the titer of the toxin in this cultivar at later growth stages represented the worst-case scenario for resistance development of MCB, since larvae of the second and third generations were exposed to sublethal concentrations of Cry1Ab toxin. Our data revealed that the variation in susceptibility to Cry1Ab for the MCB Spanish field populations analyzed in the three years was very low, with LC₅₀ values fluctuating between 12 and 30 ng Cry1Ab/cm², regardless of the region of origin, the type of maize (Bt or non-Bt) and the year. Furthermore, no significant differences were found when comparisons were made with a laboratory population (LC₅₀ values: 18-26 ng Cry1Ab/cm²) or with field populations from Greece (Bt-free areas), which displayed LC₅₀ values ranging between 22 and 27 ng Cry1Ab/cm². Standardizing bioassay protocols proved to be essential for obtaining comparable results. These findings suggest that resistant MCB populations did not evolve in those Spanish maize areas where Compa CB was largely cultivated for eight years, contradicting the expected rapid development of resistance under these unfavourable conditions. Additionally, our results can be used as baseline indices in post-market resistance monitoring programs if Bt maize is introduced in Greece. Further studies should continue, since the insights gained from a resistance monitoring program may help to enhance the durability of Bt maize.     

Occurrence,distribution, and ear damage of Helicoverpa zea (Lepidoptera: Noctuidae) in mixed plantings of non-Bt and Bt corn containing Genuity® SmartStax™ traits

Protein contamination on refuge kernels due to cross-pollination from Bt corn to non-Bt corn ears is a major concern in the use of a seed mixture refuge strategy (“RIB”) for resistance management of ear-feeding pests. In this study, occurrence, distribution, and ear damage of the corn earworm, Helicoverpa zea (Boddie), were evaluated in three planting patterns of non-Bt and Bt corn plants containing Genuity^® SmartStax™ traits. The three planting patterns were 1) pure stands of 27 Bt plants; 2) pure stands of 27 non-Bt plants; and 3) one non-Bt plant in the center surrounded by 26 Bt plants. A total of six trials were conducted in open field conditions with natural infestations in 2011 and 2012. Egg populations of H. zea were distributed randomly or uniformly, and the number of eggs laid was similar between Bt and non-Bt corn ears regardless of the planting patterns, suggesting that females of H. zea have no egg-laying preference between Bt and non-Bt plants. Bt corn plants containing Genuity^® SmartStax™ traits were equally effective in the control of H. zea in pure stands of Bt corn and “RIB” plantings. Occurrence of larvae and ear damage on Bt corn were significantly lower than on non-Bt plants and there were no significant differences between pure stands of Bt and “RIB” plantings across all trials. However, the limited numbers of live larvae in the pure stands of Bt plants were distributed non-randomly, suggesting a possibility of uneven expression of Bt proteins or elevated larval movement in the pure stands of Bt plants. Larval occurrence (3rd–5th instars) and ear damage on the refuge ears in “RIB” plantings were similar to or greater than found on ears of pure stands of non-Bt plants. However, more studies are needed to understand the effect of pollen movement on the full life cycle of H. zea before a final conclusion on the refuge function of RIB planting can be made.     

大豆和玉米加工产品中外源转基因成分检测技术的建立

采用PCR技术检测CaMV 35S启动子,并进一步通过PCR检测RoundUp Ready Soybean(RRS)和Btl76 Maximaizer的特异性DNA片段,判断大豆和玉米加工产品中是否含相应转基因成分.在1份豆粕和豆腐样品中检测到了RoundUp Ready大豆特异性的498 bp片段,而在玉米粒样品中检测到了Btl76特异性转基因成分.PCR检测的灵敏度达到0.1%,稳定性良好.结果表明,PCR技术检测外源基因是灵敏和准确的,可以广泛地应用到转基因作物及其加工产品的转基因成分检测中.