基因枪在水稻遗传转化中的应用及其转化技术的优化

1983年Zambryski等人用根瘤农杆菌介导法进行烟草基因转移,获得了世界上首例转基因植株.随后,应用DNA直接导入技术如电击法(electroporation)和PEG介导法(PEG—mediated)成功地获得了转基因水稻植株.近年来,随着基因枪技术的建立和发展,水稻遗传转化成功的报道逐年增多.目前基因枪技术在植物遗传转化中的应用超过了根瘤农杆菌介导和其它转化方法的应用.这是因为基因枪转化技术不受植物种类的限制,不需要以原生质体作为转化的受体,可以将外源基因直接导入细胞、组织或器官,因而克服了根瘤农杆菌     

根癌农杆菌转化禾谷类作物及影响其转化的因素

综述了根癌农杆菌转化禾谷类作物的研究现状,根癌农杆菌与禾谷类作物间的相互作用研究,根癌农杆菌成功转化禾谷类的例子;影响根癌农杆菌转化成功的因素,如菌株类型,感受态细胞的选择,Vir基因的活化,选择合适的转化途径等。这些将为利用根癌农杆菌介导的方法,将外源基因导入禾谷类作物提供有益的帮助。     

基因枪在植物遗传转化中的应用

基因枪技术是近年发展起来的一项新的植物遗传转化技术,本文较为系统地介绍了基因枪的类型,以及它们各自的特点和工作原理,并对基因枪在植物遗传转化中的广泛用途作了重点介绍,文中引用用参考文献七十余篇,为进一步深入了解学科的最新研究进展提供了方便。     

应用于禾谷类作物转化中的报告基因

应用于禾谷类作物转化中的报告基因邵宏波,初立业（四平师范学院生物工程研究室,吉林四平１３６０００）关键词禾谷类作物,报告基因,基因转移由于基因转移技术的发展,目前已经产生了许多的转基因禾谷类作物。为了利用基因工程方法改良禾谷类作物的品质与重要的农业性...     

农杆菌及基因枪在玉米遗传转化中的应用

目前外源基因导入玉米受体细胞的方法很多 ,最受瞩目的是农杆菌介导法和基因枪法。就农杆菌、基因枪转化玉米的基本原理、影响转化率的因素 ,以及近年来在玉米遗传转化中的最新动态进行了综述     

基因枪在植物遗传转化上的应用

把外源基因转入细胞的技术对于基因功能的研究和作物品种的遗传改良具有重要意义。随着分子遗传学、细胞生物学等基础学科及DNA重组和基因转移技术的迅猛发展,人们最终将能对进化上较为复杂的真核生物的遗传性 状进行有效的控制。     

谷类作物胚乳性状遗传控制的鉴别

谷类作物种子胚乳生状的遗传表达,可能受三倍体的胚乳基因型和／或二倍体的母体基因型控制,还可能存在细胞质效应。Ｆｏｏｌａｄ等和Ｐｏｏｎｉ等提出的分析模型,由于遗传参数的系数间存在线性相关,不能有效地区分上述遗传控制系统。本文的交配设计和相应统计方法,可对一胚乳生状是否存在胚乳基因型,母体基因型蔌细腻质效应分别作出测验,从而可确定合适遗传模型和估计有关遗传参数。这些方法也可方便地推广于鉴别非谷作物种子     

重要禾谷类粮食作物的遗传转化

就转化方法、报告基因、筛选系统和外源基因的整合与表达调控等内容 ,对近几年水稻、玉米等几种重要禾谷类粮食作物遗传转化的研究进展作了介绍和展望。     

Agrobacterium-mediated transformation of cereals: a promising approach crossing barriers

Cereal crops have been the primary targets for improvement by genetic transformation because of their worldwide importance for human consumption. For a long time, many of these important cereals were difficult to genetically engineer, mainly as a result of their inherent limitations associated with the resistance to Agrobacterium infection and their recalcitrance to in vitro regeneration. The delivery of foreign genes to rice plants via Agrobacterium tumefaciens has now become a routine technique. However, there are still serious handicaps with Agrobacterium -mediated transformation of other major cereals. In this paper, we review the pioneering efforts, existing problems and future prospects of Agrobacterium -mediated genetic transformation of major cereal crops, such as rice, maize, wheat, barley, sorghum and sugarcane.     

Rethinking sources of nitrogen to cereal crops

Understanding how to manage N inputs to identify the practices that maximize N recovery has been an organizing principle of agronomic research. Because growth in N fertilizer inputs is expected to continue in an ongoing effort to boost crop production over coming decades, understanding how to efficiently manage recovery of fertilizer N will be important going forward. Yet synthesis of published data that has traced the fate of ¹⁵N‐labeled fertilizer shows that less than half of the N taken up by crops is derived from current‐year N fertilizer. The source of the majority of N in crops is something other than current‐year fertilizer and the sources are not really known. This is true for maize (only 41% of N in crops was from current‐year N fertilizer), rice (32%), and small grains (37%). Recovery of organic fertilizer N (manure, green manure, compost, etc.) in crops is low (27%), though N recovery in subsequent years (10%) was greater than that for mineral fertilizers. Thus, while research on efficiency of N fertilizer use through improved rate, type, location, and timing is important, this research fails to directly address management of the majority of the N supplied to crops. It seems likely that the majority of non‐fertilizer N found in crops comes from turnover of soil and crop residue N. We encourage the research community to revisit the mental model that fertilizer is a replacement for N supply from turnover of soil organic N (SON) and consider a model in which N fertilizer augments ongoing SON turnover and makes an important longer term contribution to SON maintenance and turnover. Research focused on the efficient recovery of N current‐year fertilizer inputs neglects this potential role for building soil N and managing soil N turnover, which seems likely to be the most important source of crop N.     

Stem borer infestation of cereal crops in Jere,the Sudan-Sahelian savanna region of Nigeria

Field data collected during the rainy season of two years, 2010 and 2011, were used to determine the per cent plant infestation and stem borer abundance on cultivated cereal crops grown by farmers' in Jere or the Sudan-Sahelian savanna ecological region of Nigeria. Stem borers were recovered using destructive sampling. Mean total per cent plant infestation and stem borer abundance per farmers' field were significantly higher on millet (40% and 25 individuals, respectively) and sorghum (30% and 21 individuals, respectively) than on maize (19% and 13 individuals, respectively). Of the five stem borer species found in this study, Coniesta ignefusalis (Lepidoptera: Pyralidae) (3.5)/Chilo sp. nr. aleniellus (Lepidoptera: Crambidae) (2), Busseola fusca (Lepidoptera: Noctuidae) (3.1)/Sesamia calamistis (Lepidoptera: Noctuidae) (2.4) and S. calamistis (2.9), with significantly higher number of individuals per plant, tended to be more important pests on millet, sorghum and maize crops, respectively. Although, mean total per cent plant infestation and abundance of stem borers in this study were generally moderate, further studies on the effects of different types of cereals intercropping (locally practiced) on stem borer infestation and abundance should ascertain the true importance of these pest species in the Sudan-Sahelian savanna ecological region of Nigeria.     

Natural occurrence of entomophthoroid fungi (Entomophthoromycota) of aphids (Hemiptera: Aphididae) on cereal crops in Argentina

The spectrum of entomophthoroid fungal species parasitising aphids on cereal crops and a study of the phenology and prevalence of these pathogens were investigated in Argentina. The studies were conducted at six different sites cultivated with crops of Triticum aestivum (wheat), Avena sativa (oats) and Sorghum bicolor (sorghum) during two consecutive years. Entomopathogenic fungi from the new phylum Entomophthoromycota were recorded from six aphid species on cereals in Argentina: Rhopalosiphum maidis, Rhopalosiphum padi, Rhopalosiphum rufiabdominalis, Schizaphis graminum, Sitobion avenae and Sipha maydis. Three species of entomophthoroid fungi were found infecting these aphid species: Pandora neoaphidis, Zoophthora radicans (Entomophthorales: Entomophthoraceae) and Neozygites fresenii (Neozygitales: Neozygitaceae). Entomophthoroid fungal infections occurred mostly in autumn–winter seasons (March–August), and coincided with periods of high relative humidity and comparatively low temperatures. This study represents the first base‐line characterisation of entomophthoroid fungi infecting aphids on cereal crops in Argentina.     

影响基因枪法遗传转化的因素

通过对影响基因枪法遗传转化的因素探讨,以期对提高基因枪法的转化效率提供参考。     

Shoot apical meristem: A sustainable explant for genetic transformation of cereal crops

Summary Immature zygotic embryo has been the widely used explant source to develop embryogenic callus lines, cell suspensions and protoplasts for transformation of cereal crops including maize, wheat, rice, oat, barley, sorghum, and millet. However, the lack of competence of immature embryos in certain elite lines is still a barrier to rontine production of transgenic cereal crops in certain commercial cultivars. In addition, a great deal of effort is required to produce immature embryos, manipulate cultures, of immature embryos or their cell suspensions, and cryoperserve cultures for further use. In addition, undifferentiated cells may have reduced regenerability after a few months, of in vitro culture. Alternative explants and regeneration systems for efficient transformation of cereal crops are needed to avoid or reduce the above limitations. During the past decade, scientists have successfully manipulated the shoot apical meristerms from seedlings of maize oat, sorghum, millet, wheat, and barley in an effort to develop a less genetype-dependent and efficient cereal regneration system that can be maintained in vitro for long pertiods of time without the need for cryopreservation. Furthermore, apical mesistem regeneration systems were used to stably transform maize, wheat, rice, oat, barley, sorghum, and millet.     

The effect of three or five years of set-aside on the husbandry and grain yield of subsequent cereal crops in the UK

During the period 1993–1997, at six contrasting sites located throughout England, two successive cereal test crops were grown both with and without nitrogen fertiliser after three or five years of set-aside or after continuous arable cropping. Vegetation during set-aside included natural regeneration and perennial rye-grass (Lolium perenne) with or without white clover (Trifolium repens), managed by mowing on one or more occasions per year. Establishment of the successive cereal test crops after destruction of the set-aside was generally not a problem. Fertile tiller numbers were increased by inclusion of clover in the set-aside cover or application of inorganic nitrogen. The presence of couch grass (Elytrigia repens) or volunteer cereals in the set-aside covers provided alternative hosts for take-all (Gaeumanomyces graminis) and eyespot (Pseudocercosporella herpotrichoides) and take-all caused some yield reductions in following cereal crops. Management during the set-aside period significantly affected grain yields of the subsequent cereal crops in the majority of the site-year combinations. However, these effects were not as large as would be expected after traditional break crops and were frequently masked by the application of nitrogen fertiliser. Mean yields increased by 80% due to the application nitrogen at the optimum rate compared to nil nitrogen. Most of the effects of set-aside treatment on grain yield were shown to be attributable to soil mineral nitrogen content, but at some sites, infections by take-all or eyespot also accounted for some of the variation. There were no effects of pests that could be related to treatment. The presence of sown clover during the set-aside period had the most consistent effect across sites, affecting tiller populations, grain yield and grain quality of cereal crops. At some sites, establishing a sown cover during the set-aside period, or cutting the cover more than once a year, improved grain yield and quality, and reduced the incidence of some specific weeds and disease. This revised version was published online in June 2006 with corrections to the Cover Date.     

Particle bombardment and the genetic enhancement of crops: myths and realities

DNA transfer by particle bombardment makes use of physical processes to achieve the transformation of crop plants. There is no dependence on bacteria, so the limitations inherent in organisms such as Agrobacterium tumefaciens do not apply. The absence of biological constraints, at least until DNA has entered the plant cell, means that particle bombardment is a versatile and effective transformation method, not limited by cell type, species or genotype. There are no intrinsic vector requirements so transgenes of any size and arrangement can be introduced, and multiple gene cotransformation is straightforward. The perceived disadvantages of particle bombardment compared to Agrobacterium-mediated transformation, i.e. the tendency to generate large transgene arrays containing rearranged and broken transgene copies, are not borne out by the recent detailed structural analysis of transgene loci produced by each of the methods. There is also little evidence for major differences in the levels of transgene instability and silencing when these transformation methods are compared in agriculturally important cereals and legumes, and other non-model systems. Indeed, a major advantage of particle bombardment is that the delivered DNA can be manipulated to influence the quality and structure of the resultant transgene loci. This has been demonstrated in recently reported strategies that favor the recovery of transgenic plants containing intact, single-copy integration events, and demonstrating high-level transgene expression. At the current time, particle bombardment is the most efficient way to achieve plastid transformation in plants and is the only method so far used to achieve mitochondrial transformation. In this review, we discuss recent data highlighting the positive impact of particle bombardment on the genetic transformation of plants, focusing on the fate of exogenous DNA, its organization and its expression in the plant cell. We also discuss some of the most important applications of this technology including the deployment of transgenic plants under field conditions.