甘蓝转录因子BoLH27的克隆与转基因甘蓝的表型分析

bHLH转录因子对植物生长发育、形态调控有重要作用,为了研究BoLH27基因在甘蓝叶片发育及形态建成中的调控功能,本文以甘蓝品种519为材料,克隆出转录因子BoLH27基因。序列分析表明,该基因含有一个795 bp的开放阅读框,编码264个氨基酸,具有一个保守的典型bHLH结构域。以农杆菌介导的遗传转化方法将正义BoLH27基因导入甘蓝品种519中以增强表达,通过PCR筛选出9株T0代转基因单株,结合T2代单株的q RT-PCR分析表明,筛选的转基因植株内BoLH27基因的表达积累量明显高于野生型。试验基地隔离网内种植的转基因甘蓝表型明显,主要表现为莲座期茎叶间距拉长、叶柄伸长以及植株茎和叶柄显示紫色,植株叶片平展,无向上向内卷曲趋势,表明BoLH27基因可能对甘蓝叶片发育有重要的调控作用。     

国内外果蔬生物保鲜方法的研究现状与展望

概述果蔬生物保鲜方法的种类、原理和技术,同时对现阶段生物保鲜技术在果蔬贮藏保鲜中的应用进行综述,最后对生物保鲜技术存在的问题和未来研究方向进行展望.     

Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.     

小麦高分子量谷蛋白亚基研究进展

小麦是重要的粮食作物,随着人们生活水平的提高,对小麦品质提出了更高的要求,小麦高分子量谷蛋白亚基(HMW-GS)无疑对小麦品质有重大的影响。从高分子量谷蛋白亚基的命名、基因的定位、多态性和遗传以及高分子量谷蛋白亚基的结构和功能、与品质的关系、目前高分子量谷蛋白亚基分子标记的开发、转基因情况等方面进行了综述。     

蚜虫基因RNAi载体的构建及其在转基因烟草中dsRNA的表达分析

RNAi机制可以通过dsRNA诱导同源mRNA的降解，从而导致该基因转录后沉默。RNA干扰作用从发现以来就具有了利用转录后基因沉默（PTGS）来进行抗虫的一种潜在可能性。针对dsRNA的这种作用，研究中提取蚜虫RNA，克隆出蚜虫Cathepsin B基因的干扰片段并构建出RNAi载体，转入本氏烟草中，在转基因烟草体内表达dsRNA。成功构建RNAi载体后，通过农杆菌转化和叶盘法将RNAi载体导入烟草中并采用组织培养方法培养出转基因烟草，通过PCR鉴定显示15株生根的转基因烟草中14株显示阳性。提取转基因烟草的RNA进行RT-PCR和Northern表达分析显示转基因烟草植株中dsRNA表达的存在。实验结果说明了在转基因植物体内表达RNAi载体可以产生dsRNA，从而在蚜虫进食植物时达到利用dsRNA抗蚜虫的作用。     

主要农作物转基因飘流频率和距离的数据调研与分析Ⅰ.背景、调研目的及所考虑的问题

转基因飘流及其可能引起的环境和食品潜在风险是公众关注的热点之一.对2010年前主要农作物转基因飘流的数据和信息进行了调研与分析,特别是对一定允许阈值下的基因飘流距离进行了归纳.在分别报道各种作物的基因飘流数据前,对调研的背景、目的及所考虑的问题进行了讨论,基于科学分析,建议采用分类管理和阈值管理的原则来控制转基因飘流的...     

半滑舌鳎vasa调控区的克隆、表达载体构建及其驱动活性检测

为研究半滑舌鳎(Cynoglossus semilaevis)vasa(Csvasa)基因调控区的功能,在已克隆的Csvasa基因编码序列的基础上,采用基因组步移和PCR扩增的方法克隆得到Csvasa调控区,通过生物信息学方法分析vasa基因5′区,并构建了含Csvasa基因调控区的绿色荧光蛋白(GFP)表达载体(p Csvasa-GFP-T),进一步通过显微注射技术初步验证调控区的驱动活性。结果表明,通过基因组步移和PCR扩增获得Csvasa 5′区5 166 bp和3′区1 655 bp,利用在线生物信息学软件对5′区序列进行分析,发现在转录起始点上游26 bp处存在保守的TATA框,以及潜在的转录因子结合点如SRY、Oct-1、Sox-5、CREB、GATA、AP-1、C/EBP、Sp-1、c-Myc、HNF、NKX2-5、V-Myb等。通过显微注射技术,将所构建的p Csvasa-GFP-T表达载体注射于青鳉(Oryzias latipes)受精卵并进行培养观测,发现Csvasa调控区能够驱动GFP在青鳉胚胎内表达,荧光表达率为81%。将有荧光的胚胎培养为成鱼,检测外源基因的整合率为11.5%。这些结果为进一步研究半滑舌鳎原始生殖细胞(PGCs)的标记、追踪和操作研究以及半滑舌鳎的性别控制等奠定了基础。     

转兔角蛋白基因改良棉纤维品质研究

通过花粉管通道转基因技术,将E6启动子驱动的兔角蛋白基因导入高产棉花品种苏棉16号。所用转基因表达载体还含有选择标记基因NPTⅡ(卡那霉素抗性基因)及Gus报告基因。对转化体后代的检测结果表明,T1代有2.1%呈现Gus阳性,在Gus阳性株中84.6%具有卡那霉素抗性。用依据E6启动子序列和兔角蛋白基因序列设计的两对引物,对经过上述筛选的植株进行PCR检测,多次重复,最终确定3株结果稳定的转兔角蛋白基因棉株。从品质分析结果看,这3个株系成熟棉纤维的品质部分得到改良,尤其比强度有较大幅度提高,与转基因受体相比平均提高6.3cN·tex 1。