辽宁绒山羊皮肤毛囊mir-1298-5p靶基因预测及表达载体构建

探讨辽宁绒山羊皮肤毛囊不同发育时期mir-1298-5p的调控作用及其与靶基因的潜在关系,为mir-1298-5p与其靶基因对皮肤毛囊发育作用提供理论依据。以辽宁绒山羊皮肤毛囊组织为材料,通过miRNA的分离、表达、靶基因预测筛选及靶位点3'UTR表达载体构建,采用RT-PCR进行表达检测,在线靶基因预测软件预测mir-1298-5p的靶基因,并对其靶基因TGF-βR1的靶位点进行克隆测序、缺失突变。结果表明：靶基因预测发现TGF-β R1的3'UTR存在mir-1298-5p种子区结合位点,且mir-1298-5p及其靶基因TGF-β R1均在皮肤毛囊不同发育时期呈现差异性表达。mir-1298-5p对皮肤毛囊周期性发育可能起到一定的调控作用,并成功构建了TGF-βR1 3'UTR表达载体,为后期过表达转染体系的建立和功能基因的验证奠定了基础。     

亚麻转基因技术研究的进展及存在的问题

论述了20年来转基因技术在亚麻育种领域应用研究进展：(1)国内外学者成功地鉴定和克隆了亚麻目的基因;(2)进行了多种基因转化方法的研究;(3)建立了以根癌农杆菌为介导的亚麻遗传转化系统,获得了一批转基因亚麻新品系(种)。还对目前亚麻转基因研究中存在的基因转化率低、转化的外源基因可操作性差、转基因植株遗传不稳定性等问题进行了分析。     

辐射诱变在亚麻育种上的应用研究进展

20世纪20年代和40年代，在先后肯定了X射线和化学药剂有一定诱变作用之后，诱变育种工作逐渐开展起来。由于诱变育种具有提高突变率、改良品种性状、缩短育种年限的特点，因此受到国内外育种学家的广泛关注。20世纪50年代，随着原子能技术的广泛应用，采用辐射诱变比化学诱变更为普遍。1995年6月，在维也纳召开的关于应用诱发突变和分子技术改良作物的国际研讨会上确认，辐射诱变已经成为世界各国改良种质性状的有效手段，有52个国家选育出约1800个突变品种。在我国，辐射诱变的研究也取得重要进展。截至1994年底，我国在40多种植物上育成430多个优良突变品种，年种植面积900万hm^2以上，粮、油、麻等增产50亿kg，创造了巨大的经济效益。     

显性核不育亚麻在育种上的应用研究初报

利用显性核不育亚麻的育性稳定、不育株标记性状明显等遗传特点,进行了轮回选择、品种改良、培育新品种等多种利用途径的探讨。选育出产量超对照（内亚二、三号）８．１％～１８．９％的优良品种“内亚四号”;改良了“内亚二号”不抗枯萎病的缺点;创建了两个轮选群体;选育出２０个品系和一大批优异的单株材料。     

棉花转基因“喷花法”应用条件优化的研究

针对目前众多的棉花转基因方法存在转化过程复杂、成本高、效率较低等问题,实验对转基因方法一“喷花法”进行优化,明确了在棉花育种科研中应用条件,达到快速、高效转化外源基因的目的。实验选用了5个陆地棉品种(系)和2个海岛棉品种为转化受体材料,转化了2个外源基因-杭虫(Bt+CpTI)和增强纤维品质基因(NPTII-GhCAD6),并对转化时等渗液各成分的比例、转化的时间段、温度及湿度条件等因素对转化率的影响进行研究。结果表明,转化效率最高时等渗液各成分的含量:陆地棉为1/2MS+10%蔗糖+0.5g/L MES+0. Olmg/L6-BA+200u1/L表面活性剂,pH=6菌株浓度OD600=0.5;海岛棉为1/2MS+15%蔗糖+0.5g/L MES+0.01mg/L6-BA+200u1/L表面活性剂,pH=6菌株浓度OD600=0.5;转化效率最高时转化的时间段为11:00~13:30,相应的温度为22~29℃、湿度为70%~85%;转化后的T0代受体材料,首先在田间进行杭生素检测,然后在获得的阳性植株中,转杭虫基因材料用金标BT-Gry I Ab/Ac试纸条定性检测,转纤维品质基因材料做PCR电泳检测,经检测最终获得了一批转基因阳性植株。最终建立一种应用在农业育种科研中大批量、高效快速、简易实用的转基因方法。     

用磁珠富集法分离亚麻基因组微卫星分子标记

应用Dynal磁珠-生物素标记的微卫星探针(CT)₁₅与亚麻基因组DNA酶切片段杂交,捕获300~1 500 bp含有微卫星序列的DNA片段,连接到pMD18-T载体中,构建富集微卫星序列的小片段插入文库。利用接头引物和根据微卫星核心序列设计的引物VRV(CT)₁₅使用PCR方法直接对文库筛选,从422个转化子中获得了104个阳性克隆,对其进行测序分析,获得了97个微卫星序列,微卫星序列的富集效率达到22.99%,PCR扩增筛选效率93.27%。对97个微卫星序列进行比对分析,其中51个重复序列的两端序列高度相似,据其设计的特异引物对阳性克隆进行2次筛选,能淘汰相似度高的同类序列,提高筛选亚麻微卫星标记的效率。     

DTOPSIS法在亚麻新品种评价中的应用初探

为了更加准确地、有效地对亚麻新品种进行综合评价,此文以2004~2005年度云南省亚麻区域试验汇总资料为分析材料,应用DTOPSIS法对区试的10个亚麻新品种的各性状进行了综合评价,同时与传统单纯产量评价进行对比,结果表明：采用DTOPSIS法综合评价 值排序结果比单纯产量排序结果有所不同,但DTOPSIS法分析更为合理,它在全面评价亚麻品种的综合性状方面具有更好的应用价值。此外,该文还对DTOPSIS法在亚麻品种进行综合评价中有关问题进行了讨论。     

花粉管通道法在玉米转基因中的应用

对基因工程技术及花粉管通道法的发展及作用机理做了简单的介绍,概括了将外源DNA导入玉米的方式、后代变异的情况及以后的发展方向.     

亚麻新品种华亚6号及其栽培观赏应用

华亚 6 号是黑龙江省农业科学院经济作物研究所 2005 年从波兰引进种质资源材料 Pekinense 变异单株,采用系统选育法以高油、纤维、观赏性强为目标进行定向选择育成的油纤赏兼用型亚麻品种。 2019 年通过非主要农作物品种登记,登记编号： GPD 亚麻（胡麻）（ 2019 ） 230011 ,具有粗脂肪含量高、纤维产量高、高抗枯萎病且观赏性强的优点。     

花粉管通道法在玉米转基因育种中的应用

花粉管通道法是上世纪70年代提出并用于改造植物材料的基因转化方法,目前在很多农作物上有所应用。本文对该法在玉米自交系改良上的研究内容、成果进行综述,并对其今后在玉米品种改良方面的进一步应用做了展望。     

胡麻脂肪酸去饱和酶基因(fads)差异表达分析

为了了解fad基因在胡麻蒴果发育过程中对不饱和脂肪酸的调控,对高、中、低三个不同亚麻酸(C18:3)含量的胡麻品种(’CDC Gold’,‘内亚7号’,’Linola’)进行了不同时期的品质测定,以及脂肪酸去饱和酶2a基因(fad2a)、脂肪酸去饱和酶2b基因(fad2b)、脂肪酸去饱和酶2c基因(fad2c)、脂肪酸去饱和酶3a基因(fad3a)、脂肪酸去饱和酶3b基因(fad3b)的qRT-PCR定量分析。结果表明,随着蒴果成熟,可溶性糖含量呈降低趋势,粗脂肪与粗蛋白不断积累,且差异显著(p<0.05)。fad2a基因、fad3a基因以及fad3b基因在各个时期中的表达符合正态分布。以0 d的蒴果为对照,在胡麻种子形成过程中,‘内亚7号’的三个基因在5 d和15 d的表达量迅速增加,到30 d时急剧减少,15 d的fad2a基因表达量为5.23倍,fad3a基因表达量是fad2a基因表达量的14.52倍,fad3b基因的表达量是fad2a基因表达量的16.14倍,表明这三个基因参与不饱和脂肪酸积累过程。在低亚麻酸含量品种‘Linola’30 d中,fad3a基因是fad2a基因表达量的52.71倍,fad3b呈下调趋势;在高亚麻酸含量品种‘CDCGold’30d中,fad3a基因与fad2a基因的表达量均呈下调趋势,fad3b的表达量为3.92倍;在中等亚麻酸含量品种‘内亚7号’中,fad2a基因表达量降低了0.31倍,fad3a基因是fad3b基因表达量的1.87倍。fad2b基因、fad2c基因熔解曲线不稳定,峰值低,可以在后续试验中继续探索。     

用in planta方法转化甘蓝型油菜

用in planta方法进行遗传转化极大地促进了模式植物拟南芥的功能基因组研究,但在农作物中仍少有in planta的成功报道。我们采用in planta方法,用含有质粒pSG529、pHTT613和ApSCO的农杆菌菌株转化中油821、湘油13、宁RS-1、S2501-1、华双3号等甘蓝型油菜品种。在适宜的条件下,将根癌农杆菌菌液浸染甘蓝型油菜花序10 d至15 d     

新疆地方种亚麻农艺性状多元统计分析

为了新疆亚麻地方种的选育和改良提供理论依据,本研究采用对来自新疆各地的亚麻地方种性状的多元统计分析,包括变异系数、主成分分析、相关性分析和聚类分析。结果表明,新疆亚麻地方种资源存在较大差异,多样性比较丰富。主成分分析中,株高会影响构成亚麻产量的因子(蒴果数,果粒数,每株粒重和千粒重),亚麻的分枝数和株高都不能过多或过高,这不仅会影响工艺长度还会影响亚麻产量因子而且发现株高、生育期、分枝数、工艺长度、每株粒重、分枝数、千粒重和果粒数的相关程度较高。因此,在新疆亚麻地方种在选育时,可作为理论性数据参考,为亚麻品种选育和亲本的选择提供一定基础。     

170份亚麻种质资源主要农艺性状分析

对来自五大洲33个国家的170份亚麻种质资源的9个农艺性状进行了分析。结果表明,参试资源的农艺性状差异较大,变异丰富。各性状间相互影响,有10对性状呈极显著相关,其中,株高、分茎数、主茎分枝数、单株果数均与单株产量呈极显著正相关,作用效应为单株果数>主茎分枝数>分茎数>株高。主成分分析表明前3个主成分的累积贡献率为74.705%。将170份种质资源划分为5类,各个类群之间差异明显。对33个国家类群进行聚类分析,分为3个组群,地理来源相同的品种被归为不同类群,洲际之间的种质资源在组群中相互渗透。     

Comparison of European Fibre Flax (Linum usitatissimum) Cultivars under Eastern Canadian Growing Conditions

Renewed interest in natural fibres, decreasing subsidies to European producers, and high production costs have led the move to reintroduce fibre flax (Linum usitatissimum L.) production into eastern Canada. Research was conducted at the Macdonald Campus of McGill University, Québec, Canada in 1998 and 1999 and at Winchester and Kemptville, Ontario, Canada in 1998, to assess the performance of seven European fibre flax cultivars. Parameters evaluated included plant density, branching ratio, stem diameter, fresh and dry biomass, and mean harvest height. Data from all sites and years were subjected to a pooled anova where appropriate. The cultivar main effect was detected for all parameters measured, with the exception of mean fresh weight and mean height at harvest. There was also a site main effect for all parameters except for branching ratio. A cultivar–site interaction was found for all parameters except for mean stem diameter and mean dry weight. Results indicate a strong potential for producing fibre flax in eastern Canada using currently available European cultivars.     

Selection Criteria for Short-fibre Flax

The industrial utilization of short fibres of Linum usitatissimum L. is advantageous for the environment and conserves natural resources due to savings in the production and waste management process. In 1990 a project was started to breed flax for short fibres in combination with oil production. In field experiments 22 (1990) and 20 (1991) varieties and progenies of earlier crosses, respectively, were tested. Phenotypic variation and linear correlations of parameters characterizing yield and its stability were examined with regard to their suitability as selection criteria. The results were as follows: A total harvest of 3 t/ha of short fibre and oil was produced, most of it fibre. The cross progenies tested in 1991 showed a high genetic potential for the combined use of fibre and oil, but their maturity needs to be synchronized better. Simultaneous selection of short-fibre flax genotypes for fibre content and seed yield on the one hand, or for oil content and fibre yield on the other, seems to be practicable. The plants should not exceed I m in height. Thousand-seed weight and oil content were not correlated in the progenies, thus making it possible to select for small-seeded genotypes.     

Evaluation of Fibre Flax (Linum usitatissimum L.) Performance under Minimal and Zero Tillage in Eastern Canada

The feasibility of producing fibre flax in minimum tillage or zero tillage (ZT) systems was investigated. The results were variable between the sites, which differed in soil type and previous cropping history. Tillage regime had no impact on fibre flax phenological development including number of days to emergence, days to flowering and days to harvest at either site. However, tillage regime had a significant effect on mean stem diameter, dry matter content and plant height at all three sampling dates at the sandy loam site, and a significant effect on mid‐season plant height at the clay site. At the sandy loam site, ZT plots had the highest populations of plants with the finest stem diameters, the lowest branching ratios, but the shortest plants by the end of the season. Overall, plant densities were greater, stem diameters thinner, and biomass production higher in the heavier soil of the clay site. These results indicate that under the growing conditions present in 1998, fibre flax can successfully be grown in minimum or ZT systems on different soil types in Eastern Canada. Moreover, the growth of fibre flax in these systems does not compromise the proportion of tall plants having thin stems and minimal branching, a critical quality parameter for fibre flax production.     

生物技术在纤维亚麻育种中的应用

亚麻纤维在农业、医疗、工业及航天业得到广泛应用。随着分子生物学技术与基因工程技术的迅猛发展,育种方法的研究成为培育新亚麻植物的关键,为了加快育种效率,提高亚麻纤维的质量与产量,综述了基因工程技术、组织培养技术和分子标记技术在亚麻育种中的应用,重点阐述了基因工程在亚麻质量和品性提升中应用以及原生质体融合与培养技术在亚麻育种中应用的最新研究进展,为亚麻种质资源的研究与发展提供依据。     

Influence of Seeding Depth and Seedbed Preparation on Establishment, Growth and Yield of Fibre Flax (Linum usitatissimum L.) in Eastern Canada

Research was conducted at the Macdonald Campus of McGill University (Québec, Canada) at three sites in 1997 and one site in 1998 to determine the effects and interactions of seeding depth (0, 1, 2, 4 or 6 cm) and seedbed preparation (i.e. soil rolling): none, rolling before or rolling after seeding on fibre flax (cv. Ariane) establishment, growth and yield. Seedbed preparation had little impact on the parameters measured while seeding depth had a variable effect on plant density, plant height, stem diameter and retted straw yield. Seeding depths of 1–4 cm provided consistently good establishment, growth and yield results. In 1997, there was an interaction between seeding depth and seedbed preparation on plant height, branching ratio and retted straw yield, although results were generally variable and tended to be site‐specific. In 1998, there was an interaction between seeding depth and seedbed preparation on plant height and stem diameter prior to harvest, with the results varying for all seeding depth‐seedbed preparation treatment combinations except for the 2‐cm depth treatment. Rolling of the seedbed before seeding on lighter soils and at a depth of 2 cm on most soils can improve establishment, growth and yields of fibre flax under eastern Canadian growing conditions.     

基因工程技术在亚麻抗病育种中的应用研究进展

本文论述了20年来基因工程技术在亚麻抗病育种领域的应用研究进展：（1）亚麻抗病基因的克隆及分子标记。（2）亚麻转抗病基因的研究进展。（3）亚麻抗病转基因研究存在的问题。