Risk Assessment and Biosafety Studies of Transgenic Bt Rice （Oryza sativa L.）

Transgenic crops having alien genes from different sources are getting popularity. A Rice （Oryza sativa L.） containing gene from Bacillus thuringiensis, a soil bacterium, was assessed to study the potential risks of transgenic plants on environment. The crop was found resistant to target insect pests. Rice variety Basmati-370 transformed with two insecticidal genes, crylAc and cry2A, was grown under field conditions for several years. Data were collected at different stages of the plant growth. Fate of Cry protein in soil, effect of Bt protein on non-target insects, risks of vertical and horizontal gene flow were evaluated. No potential hazard was found at all levels. Bt protein was unstable and degraded significantly in soil within 30 days after harvesting the crop. No harmful effects were found on non-target insects （insects other than order lepidoptera）. Maximum gene flow of 0.02% was observed at close spacing and no evidence of horizontal gene transfer to Rhizobium spp. was found. In conclusion, the transgenic rice plants transformed with Bt genes have no harmful effects on the environment.     

Effects of Gradual Water Deficit Stress on Phenological and Morphological Traits in Chickpea （Cicer arietinum L.）

The aims of this study were to investigate the effect of gradual water deficit stress on some phonological and morphological traits and grain yield of desi and kabuli chickpea cultivars. This study was carried out in 2007 and 2008, to evaluate responses of three chickpea cultivars （Hashem and Arman from kabuli and Pirooz from desi type） under well watering （I1： 70mm evaporation from class A pan）, gradual water deficit （12 and 13： 70→90→ 110→130 and 70→100→130mm evaporation from class A pan, respectively） and severe water stress （14： 130mm evaporation from class A pan）. Result showed that days to flowering and plant height were decreased, as water limitation increased. This reduction was significant under gradual water stress （I2 and I3） and Severe water deficit （I4）, compared with control （I1）. There were no significant differences in grain filling period and grain yield among I~, I2 and I3 irrigation treatments. No significant differences in days to physiologic maturity and number of sub branches were recorded among irrigation treatments. Interactions of year×cultivar for days to physiologic maturity, grain filling period and grain yield （P≤0.01） and for days to flowering and plant height （P≤0.05） were significant. The superiority of Arman in producing comparatively greater grain yield could be attributed to higher grain filling period of this cultivar in both years.     

灰色理论在肇庆地区干旱预测中的应用

采用1979~2008年肇庆地区近30年的降水资料,根据《干旱评估标准》,确定肇庆干旱的灾变数据序列,建立GM（1,1）模型,采用最小二乘法求解参数a、b,得到肇庆地区GM（1,1）数列预报模型。经后验差方法检验,预测精度为＂好＂等级,故可用该模型对肇庆地区未来20年可能发生干旱的年份进行预测。预测结果显示,肇庆地区将在2009~2010、2013~2014、2018~2019、2022~2023、2028~2029年期间发生干旱。     

长白柳种质试管保存研究

[目的]筛选出长白柳嫩茎离体培养各阶段的培养基。[方法]以长白柳嫩茎基部直接再生形成的丛生芽团为试材,利用＂延缓生长＂的方法在试管内对长白柳进行种质保存,应用均匀设计法探讨了影响长白柳丛生芽团试管保存的主要因子和水平,筛选最适于长白柳丛生芽团试管保存的培养基。[结果]长白柳最适宜的丛生芽团试管保存培养基为：N-68＋根皮苷3.20 mg/L＋KT0.70 mg/L,保存43个月生长率仅为0.97%。对保存前后的芽苗生根情况对比结果表明,保存后的芽苗生根速度快,生根率达99.6%以上,较未保存前的生根率99.0%高。[结论]在试管内采取＂延缓生长＂法保存长白柳丛生芽团是切实可行的新方法。     

苏云金芽孢杆菌cry1Ac28基因的克隆及原核表达

研究旨在克隆苏云金芽胞杆菌(Bacillus thuringiensis,Bt)Q-12的cry1Ac28基因并在原核载体中表达。应用PCR-RFLP法鉴定出Bt Q-12中含有cry1Ac基因,根据已知的cry1Ac全长基因序列设计特异引物,以Bt Q-12基因组DNA为模板扩增cry1Ac全长基因,与大肠杆菌(Escherichia coli)表达载体pEB相连接获得含有cry1Ac全长基因的重组质粒pEB-cry1Ac,经过序列分析表明,该基因编码区为3 537 bp,编码1 178个氨基酸,分子质量为133.3176 ku,pI为4.885,为弱酸性蛋白质,亮氨酸(Leu)、丝氨酸(Ser)、谷氨酸(Glu)三种氨基酸含量最高,分别为8.06%、7.80%、7.72%,该基因在大肠杆菌BL21(DE3)菌株能够正常表达133.3176 ku蛋白。该基因核苷酸序列已在GenBank中登录,登录号为FJ610439,并由Btδ-内毒素基因国际命名委员会正式命名为cry1Ac28。为进一步研究cry1Ac28蛋白功能和活性打下了良好的基础。     

猪抗酶基因1(AZ1)5’调控区单核甘酸多态性与屠宰性状的相关性分析

通过PCR-RFLP方法检测了长白与蓝塘猪F2代群体抗酶基因1(AZ1)5’调控区-792～-433单核苷酸多态性,共获得3个SNP:-731C＞T,-584A＞C和-476G＞A.-713位点处T为优势等位基因,频率为0.777,优势基因型TT频率为0.598;-584位点处A为优势等位基因,频率为0.769,优势基...     

水稻密码子优化的cry1Ca 基因在大肠杆菌中表达、纯化及抗体制备

将水稻偏好密码子优化并人工合成抗虫基因cry1Ca通过限制性内切酶酶切的方法构建原核表达载体pET-28bca,成功表达了带6个组氨酸标签的His-cry1ca融合蛋白,其表达量约占菌体总蛋白的24%,表达的融合蛋白主要以包涵体的形式存在.对包涵体蛋白进行可溶性处理,亲和纯化出带6个组氨酸标签的融合蛋白,将其作为抗原免...     

黑曲霉糖化酶基因的克隆及其在毕赤酵母X33中的表达

[目的]以毕赤酵母（Pichia pastoris）X33为宿主菌高效表达黑曲霉（Aspergillus niger）糖化酶,为进一步扩大糖化酶在工业上的应用奠定基础。[方法]利用RT-PCR技术,提取黑曲霉总RNA进行反转录,以得到的cDNA为模板,根据NCBI数据库中A.nigerCBS513.88糖化酶的cDNA序列（glaA）设计引物,通过PCR扩增得到去除天然信号肽的糖化酶成熟肽编码基因glaAm,并将其克隆到pUC19载体中,序列分析表明,glaAm的开放阅读框由1 879个核苷酸组成,编码625个氨基酸。以此片段构建了pFLDα-glaAm重组表达载体,经NsiI线性化后,电击转化毕赤酵母X-33。摇瓶培养中通过添加终浓度为0.5%的甲醇诱导糖化酶的分泌。[结果]SDS-PAGE和Starch-PAGE显示,糖化酶得到正确的分泌表达,且具有较高的生物活性,发酵上清液中酶活最高达到380.78 U/ml（发酵液）。重组糖化酶的最适反应温度和最适pH分别为6065℃和4.0,在pH2.55.5范围内均保持90%以上的酶活力。该酶具有较高的热稳定性,pH4.0条件下,该酶在50℃下稳定;60℃处理60 min,仍能保持90%以上的酶活力;65℃下的半衰期约为44 min。[结论]黑曲霉糖化酶基因在毕赤酵母X33中得到了异源高效表达。     

淹水胁迫对菘蓝苗期乙醇脱氢酶与过氧化物酶活性的影响

目的:研究淹水胁迫下菘蓝中乙醇脱氢酶(ADH)和过氧化物酶(Peroxidase,POD)活性的变化,以期为进一步研究菘蓝抗涝害的机理提供参考.方法:应用NAD还原法测定淹水胁迫下菘蓝中乙醇脱氢酶活性的变化;采用愈创木酚比色法测定过氧化物酶活性.结果:淹水胁迫下乙醇脱氢酶活性被诱导,淹水处理不同时间乙醇脱氢酶活性均高于...     

Construction of Tobacco Bax lnhibitor-1 ihpRNA Gene Silencing Vector and Transformation into Agrobacterium tumefaciens EHA105

The plasmid pGSA1285 was first modified by substituting its GUS sequence with the Chalcone synthase intron fragment from vector pFGC5941 to get the plant silencing expression vector that contained Kanamycin resistance site and was named as pGSA2285. Using PCR-based amplification, two different restriction sites at both ends of tobacco Bax inhibitor-1 (NtBI-1) gene were created, respectively, which made the construction of ihpRNA gene silencing vector more efficiently. Then, NtBI-1 genes were inserted into Multiple Cloning Site (MCS) of pGSA2285 respectively to form Bax inhibitor-1 ihpRNA gene silencing vector, named as pGSA4285, containing sense and anti-sense BI-1 sequence which was spliced by chalcone synthase intron. Combined PCR identification and enzyme restriction analyses, the results showed that Bax inhibitor-1 ihpRNA gene silencing vector had been constructed and transferred into Agrobacterium tumefaciens EHA105 successfully, which laid a foundation for the further study on the function of BI-1 in plant PCD regulation.