β-伴大豆球蛋白α'亚基和β亚基基因双价RNAi表达载体抑制大豆抗原蛋白的表达

为幼龄动物提供优质的大豆饲料,同时有效抑制α'亚基和β亚基基因表达量,根据dsRNAi原理,以α'亚基基因RNAi重组植物表达载体p CAMBIA3301-PFNZ-BADH为基础,构建以BADH安全标记为筛选标记的α'亚基和β亚基基因双价RNAi植物表达载体。用根癌农杆菌介导法转化大豆子叶节,并对再生植株进行分子生物学检测。结果表明:成功构建β-伴大豆球蛋白α'亚基和β亚基基因双价RNAi表达载体,并获得大豆转基因植株。T1转基因大豆植株经PCR检测得到11株阳性转化株,经Southern杂交检测证明构建的双价植物表达载体已经以1~2个拷贝整合到大豆基因组中,实时荧光定量PCR检测表明β-伴大豆球蛋白α'亚基和β亚基基因的表达被明显抑制。     

农杆菌介导的大豆植株整体转化

农杆菌介导的植物整体转化法主要以植物的分生组织和生殖器官作为外源基因导人的受体,通过真空渗透法、浸蘸法及注射法等方法使农杆菌与受体材料接触,以完成可遗传细胞的转化,通过抗生素筛选和分子检测鉴定转基因植株后代.以大豆幼苗的顶端生长点和叶腋生长点为靶点,通过农杆菌介导的整体转化法--注射法进行转录因子DREB1C基因的遗传转化,最终获得6株T0代PCR阳性转基因植株,转化率为9.2%.T1代植株的PCR、Southern blot和RT-PCR鉴定结果表明获得1株阳性植侏,DREB1C基因以单拷贝形式整合于大豆基因组中,在200 mmol·L-1NaCl胁迫条件下在转录水平得到成功表达.该方法克服了大豆组织培养再生难、转化率低的缺点,是一种高频、简单、快捷的非组培遗传转化途径.     

农杆菌介导法将Bt(cryⅠA)基因导入大豆的研究

构建含有Bt(cry Ⅰ A)基因的植物表达载体pCAMBIA3300-Bt,以大豆子叶节为受体,通过农杆菌介导法将Bt基因导人大豆品种黑农37中,获得转基因植株.并进行人豆的再生和遗传转化系统优化的研究,以获得较高的转化率.结果表明:在6-BA浓度为1.7 mg·L-1时,丛生芽分化率最高,确定该品种大豆在从生芽分化阶段的草铵膦筛选浓度为3.5 mg·L-1获得转化质粒pCAMBIA3300-Bt的转基因植株,其中T1代PCR阳性植株19株.采用real-time PCR的方法对T1代抗性植株进行Bt基因的转录水平的分析,初步证明Bt基因已整合到受体大豆的基因组内.     

转cry 1Iem基因大豆的培育及抗虫性检测

以大豆子叶节为外植体,应用农杆菌介导法,将抗虫(cry1Iem)基因转化大豆.筛选标记为bar基因.经Glufasinate筛选,获得大量抗性植株.对转基因T_0、T_1、T_2代植株进行PCR检测,初步证明cry1Iem基因已经整合到大豆基因组中.对T2代PCR阳性植株幼嫩豆荚,采用圆盘分隔法接人初孵幼虫,进行初步的抗虫性检测,得到1株具有明显抗虫效果和7株抗虫效果较好的转基因植株.     

CryIA(c)基因植物表达载体的构建及转基因甘蔗的获得

构建了玉米UBI启动子驱动Bt CryIA(c)基因的高效植物表达载体pUBTC,通过农杆菌介导法导入甘蔗愈伤组织,经过除草剂PPT 3次连续筛选,获得89株抗性植株,经过PCR扩增检测,得到71株阳性植株,对其中7株进行RT-PCR检测,证明外源基因已经成功的整合到甘蔗基因组中,并得到有效的表达。     

Cry1Ab基因转化甘蔗及转基因抗虫植株的获得

构建玉米UBI启动子驱动Bt Cry1Ab基因的高效植物表达载体pUBTB，通过农杆菌介导法导入甘蔗愈伤组织，经过除草剂PPT三次连续筛选，共获得67株抗性植株，经PCR扩增检测，得到22株阳性植株，并对其进行RT-PCR检测，证明外源基因已经成功整合到其中的15株甘蔗基因组中，且得到了有效的表达。     

转酸性蛋白酶pepB基因玉米的分子鉴定与农艺性状分析

通过分子生物学技术把酸性蛋白酶pepB基因转入受体玉米自交系基因组中,培育转酸性蛋白酶pepB基因玉米新品系。以耐盐基因badh作为筛选标记性基因,经过抗性筛选,对T1、T2、T3代转基因植株进行PCR检测,得到14个T1代转基因阳性植株,32个T2代转基因阳性植株和27个T3代转基因阳性植株。Southern杂交结果表明,外源酸性蛋白酶基因已经整合进玉米基因组中。RT-PCR结果表明,酸性蛋白酶基因在受体玉米中获得表达,获得外源酸性蛋白酶pepB基因遗传表达的T3代转基因玉米株系。通过对T2、T3代转基因玉米各品系农艺性状的分析结果表明,转基因玉米各品系在株高、茎粗、穗长等农艺性状上与受体亲本没有差异,但生育期均有不同程度的缩短。     

农杆菌介导法将Bt（cryI4）基因导入大豆的研究

构建含有Bt（cryIA）基因的植物表达载体pCAMBIA3300-Bt，以大豆子叶节为受体，通过农杆菌介导法将Bt基因导入大豆品种黑农37中，获得转基因植株。并进行大豆的再生和遗传转化系统优化的研究，以获得较高的转化率。结果表明：在6-BA浓度为1．7mg·L^-1时，丛生芽分化率最高。确定该品种大豆在丛生芽分化阶段的草铵膦筛选浓度为3．5mg·L^-1。获得转化质粒pCAMBIA3300-Bt的转基因植株，其中T1代PCR阳性植株19株。采用real—timePCR的方法对T4代抗性植株进行助基因的转录水平的分析，初步证明成基因已整合到受体大豆的基因组内。     

花粉管通道法将Bt-CPTI双价抗虫基因转入玉米自交系的研究

利用花粉管通道法将Bt-CPTI双价抗虫基因对优良玉米自交系进行遗传转化。采用组培和盆栽两种方法对大量t0代转基因种子进行卡那霉素抗性筛选,获得的卡那抗性植株进行PCR及PCR-Southern检测,初步证明外源目的基因已整合到玉米基因组中,PCR-Southern阳性率达到2.19%。     

未成熟种子子叶节转化体系优化及转EPSPS/GAT基因大豆新材料创制

以丛生芽率为指标,将农杆菌介导的大豆未成熟种子子叶节转化与成熟种子转化体系进行比较,明确了诱导过程中草甘膦筛选浓度、未成熟种子生理状态,改进大豆外植体的获得方式并减少了组培环节,建立了未成熟种子子叶节转化体系。利用Jack黄熟后期的种子作为受体材料,在15 mg·L-1的草甘膦筛选浓度下获得了9株PCR检测阳性植株并且生长正常可育,T0代经测序及Southern Blot分析,进一步证明外源基因已整合进大豆基因组中,其中2个株系的T1代植株经PCR检测符合3∶1的分离比,且表型鉴定筛选出抗性转基因植株,为抗草甘膦转基因大豆育种提供了新材料。     

Sorghum and millet phenols and antioxidants

Sorghum is a good source of phenolic compounds with a variety of genetically dependent types and levels including phenolic acids, flavonoids, and condensed tannins. Most sorghums do not contain condensed tannins, but all contain phenolic acids. Pigmented sorghums contain unique anthocyanins that could be potential food colorants. Some sorghums have a prominent pigmented testa that contains condensed tannins composed of flavan-3-ols with variable length. Flavan-3-ols of up to 8–10 units have been separated and quantitatively analyzed. These tannin sorghums are excellent antioxidants, which slow hydrolysis in foods, produce naturally dark-colored products and increase the dietary fiber levels of food products. Sorghums have high concentration of 3-deoxyanthocyanins (i.e. luteolinidin and apigenidin) that give stable pigments at high pH. Pigmented and tannin sorghum varieties have high antioxidant levels that are comparable to fruits and vegetables. Finger millet has tannins in some varieties that contain a red testa. There are limited data on the phenolic compounds in millets; only phenolic acids and flavones have been identified.     

Tuber and starch quality of wild and cultivated potato species and cultivars

Summary Between 1993 and 1998 205 different potato cultivars and 1220 accessions/genotypes of wild and cultivated potato species from the IPK Genebank Gatersleben were evaluated. Parameters interesting for starch isolation and especially for the use of starch were determined. Altogether, there was a higher variability in wild potato species than in cultivated potatoes for all characteristics investigated: dry matter content, starch content, protein content, amylose content and mean particle diameter of starch granules.     

种子加工、检验理论与技术现状及思考

种子加工及检验是种子向市场流通的关键，是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实“育繁推一体化”种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中，受制于行业发展水平，种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程，提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要，在做好品种优质化繁育的基础上，分析种子加工及检验理论与技术的新方向和新需求，加强种业应用性研究，建立先进的种子质量检验体系，研制适合现代种业发展需求的种子加工设备，打造种子加工产业标准化生产体系，完善种业科学技术学科建设，促进种业产业持续健康发展。     

Present Situation and Consideration of Seed Processing and Testing Theory and Technology

Seed processing and testing hold the key to the market circulation of seeds, and are an indispensable link in commercial breeding to endow seeds with commodity attributes. The research of seed processing and testing theory and technology is an important link in improving the seed industry chain, realizing large-scale commercial breeding and consolidating the seed industry science of ‘integration of breeding, propagation and promotion’. The scientific and technological system of seed industry in China is being formed and perfected, which is subject to the development level of the industry. The theory and technology of seed processing and testing are relatively weak. We reviewed the development of seed processing and testing theory and technology, and put forward strategies and suggestions to improve the sound development of China’s seed industry. In order to meet the needs of seed industry development and on the basis of high quality breeding of varieties, the new directions and demands of seed processing and testing theory and technology were analyzed. We will work to strengthen the applied research of seed industry, establish advanced seed quality inspection system, improve seed processing equipment suitable for the development needs of modern seed industry, establish standardized production system of seed processing industry, develop the scientific and technological disciplines of seed industry, and promote the sustainable and healthy development of seed industry.     

种子加工、检验理论与技术现状及思考

种子加工及检验是种子向市场流通的关键,是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实"育繁推一体化"种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中,受制于行业发展水平,种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程,提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要,在做好品种优质化繁育的基础上,分析种子加工及检验理论与技术的新方向和新需求,加强种业应用性研究,建立先进的种子质量检验体系,研制适合现代种业发展需求的种子加工设备,打造种子加工产业标准化生产体系,完善种业科学技术学科建设,促进种业产业持续健康发展。     

芸薹属多倍体与二倍体作物 花柱及角果的发育差异

通过比较分析芸薹属多倍体复合种与其祖先二倍体基本种在花柱、柱头大小、角果长度、宽度、种子直径 及种子干重变化上的异同,发现多倍体复合种在这些发育性状上表现出处于两个祖先二倍体亲本之间,或只偏向 于其中一个祖先亲本。同一个多倍体品种在不同发育性状上具有不同的表现,即在某一发育性状上偏向于其祖先 亲本之一,而在另一发育性状上偏向其另一祖先亲本。     

Effects of Organic and Chemical Fertilizers and Arbuscular Mycorrhizal Inoculation on Growth and Quality of Cucumber and Lettuce

Summary

Arbuscular mycorrhizae were inoculated into phosphorus-deficient soil fertilized with either organic or chemical fertilizer with cucumber (Cucumis sativus L.) as the first crop and lettuce (Lactu-ca sativa L.) as the second crop but without additional fertilization and AM inoculation. AM increased dry matter and fruit yield of cucumber significantly in the unfertilized, organic-fertilized and P-deficient plants compared with the fully chemical-fertilized plants. AM inoculation increased the available phosphorus in plant and soil by around 30% for all treatments except for those chemically-fertilized. The rate of AM infection did not differ significantly among the fertilization treatments, but the infection intensity was higher in unfertilized, organic-fertilized and phosphorus-deficient treatments than chemical-fertilized treatment. The residual effects of AM-inoculated to cucumber were evident for lettuce in all pre-treatments that were unfertilized and un-inoculated for the second cropping. Without P-fertilization, neither crop could grow optimally even when the soil was inoculated with AM, suggesting that AM could not serve as a substitute for phosphorus fertilizer. However, the other beneficial effects of AM on crop growth and yield could not be fulfilled with phosphorus fertilizer.     

Effects of crop density and tillage system on grain yield and N uptake from soil and atmosphere of sole and intercropped pea and oat

Pea (Pisum sativum L.) and oat (Avena sativa L.) were grown as sole and mixed crops in various densities under two different tillage systems on a loess soil near Göttingen/Germany in a 2-year field experiment (2002/2003). In the conventional tillage system a mouldboard plough (CT) was used and in the minimum tillage system a rotary harrow (MT) was employed. The effect of crop density and tillage system on the grain dry matter and grain N yields, N₂ fixation and soil N uptake were determined to address the following questions: (i) which mixture compositions exhibit the highest grain yields compared to the sole crops, (ii) which mixture compositions also fix a high level of N₂ and leave low levels of residual inorganic soil N after harvest, and (iii) whether the intercrop advantage is influenced by the tillage system. For (i) the result in 2002 showed that the highest grain yields of both sole cropped pea and oat and intercropped pea and oat were achieved at the highest densities. In 2003, when the inorganic soil N content was higher and weather conditions were warmer and drier, grain yields were significantly higher than in 2002, but sole as well as intercropped pea and oat gave their highest grain yields at lower densities. For both years and tillage systems, the highest intercrop advantages were achieved in mixtures with densities above the optimal sole crop densities. The result for (ii) was that a distinctly higher proportion of nitrogen was derived from the atmosphere (Ndfa) by intercropped pea than by sole cropped pea. However, the uptake of soil N by intercropped pea and oat was not reduced in comparison with that of sole cropped oat as the decrease in the uptake of N from the soil by oat at lower oat densities in the mixture was compensated for by the soil N uptake of pea. Additionally, the N_min-N content of the soil following the mixtures and sole cropped oat did not differ, especially in the deeper soil layers because oat in mixture was forced to take up more soil N from deeper layers. Therefore, the risk of soil N losses through leaching after mixtures was lower compared to sole cropped pea. The tillage system (iii) had no significant influence on grain yield and soil N uptake, but N₂ fixation and the competitive ability of intercropped pea were higher under CT than with MT. An additional result was that intercropping led to a significantly increased grain N content of both pea and oat compared to the sole crops. The increase in grain N content from sole to intercrop was from 3.30 to 3.42% for pea and from 1.73 to 1.96% for oat as a mean for both years and tillage systems. The present study confirms that growing pea and oat as intercrops highlights potential economic and environmental benefits which still need to be understood in more detail in order to exploit intercropping to a greater extent.     

云南咖啡主要病虫害及防治调查研究

对云南保山、德宏、普洱等地咖啡病虫害开展调查研究,结果表明：咖啡锈病(Hemileia vastatrix Berk et Br.)、炭疽病(Colletotrichum coffeanum Noack)、褐斑病(Cercospora coffeicola Berk. et Cooke)、幼苗立枯病(Rhizoctonia solani Kuhn.)、旋皮天牛(Acalolepta cervina Hope)、灭字虎天牛(Xylo     

我国麻类标准现状与分析

介绍了我国麻类标准的现状。通过分析,指出了存在的问题,主要表现在标准体系不完善、标准更新不及时,协调性差及贯彻执行不力等。提出了我国麻类标准应采取的对策。