安祖花Arizona品种遗传转化体系的优化及矮牵牛查尔酮合成酶基因(PhCHS)的导入

安祖花(Anthurium andraeanum)是近年来非常流行的年宵花卉之一。为了优化安祖花的遗传转化体系,本研究以安祖花Arizona品种组培苗的叶片为外植体,将构建好的重组质粒p SN1301-HYG-Ph CHS在根癌农杆菌(Agrobacterium tumefaciens)GV3101介导下转化安祖花。优化后的体系为:首先将组培苗幼嫩叶片切块后置于1/3 MS预培养基(附加200 mg/L NH4NO3,0.5mg/L BA,0.05 mg/L 2,4-D,5g/L琼脂,30g/L蔗糖)中预培育3 d,然后用根癌农杆菌(菌液OD600=0.5)侵染10 min,将侵染过的叶片接种于MS培养基,28℃黑暗条件下共培养3 d,之后转移到附加有500 mg/L羧苄青霉素和30 mg/L潮霉素的愈伤诱导培养基中进行筛选。结果表明,抗性愈伤组织形成率可达到5%~6%。对体系优化过程中获得的14株抗性转基因植株进行PCR检测,其中9株为阳性,证实矮牵牛查尔酮合成酶基因(Petunia hybrida chalcone synthase gene,PhCHS)已经转入安祖花Arizona中。该研究结果为利用转基因方法改良安祖花品种提供了技术保障。     

谷子显性恢复基因的AFLP分析

赤峰显性核不育谷子是在谷子中首次发现的核不育材料,该材料的育性受2对核显性基因互作控制,一对是显性核不育基因Msch,另一对是显性上位育性恢复基因Rf。两者共同存在时显性上位育性恢复基因Rf能抑制显性核不育基因Msch的表达,从而表现可育。利用已构建的不育基因Msch的上位育性恢复基因Rf的近等基因系(NILs)为材料,通过对300对AFLP引物组合进行筛选,找到了与显性上位育性恢复基因Rf紧密连锁的2个AFLP标记(E15/M52和E20/M41),与不育基因的遗传距离分别是7.0c M和12.7 c M,而且位于不育基因的同一侧,标记间相距5.7 c M。     

笼养条件下花尾榛鸡的遗传多样性分析

[目的]探讨笼养花尾榛鸡的遗传分化。[方法]采用PCR和DNA直接测序技术测定线粒体细胞色素Cyt b基因全序列,结合Gene Bank收录的其他松鸡科同源序列,利用生物学软件进行分析。[结果]15只花尾榛鸡样本Cyt b基因序列中,共检测到核苷酸多态位点12个,具有6种单倍型,笼养吉林花尾榛鸡遗传多样性水平较低。系统分析结果显示,吉林花尾榛鸡与松鸡科榛鸡属亲缘关系最近,与北美榛鸡属亲缘关系较远。[结论]花尾榛鸡属于松鸡科榛鸡属,不应并入北美榛鸡属。     

SSR标记技术及其在杨梅上的应用

从SSR位点类型、SSR位点遗传特性、SSR标记技术原理与特点及SSR标记开发等方面对SSR标记技术进行概述;并就SSR标记技术在杨梅种质资源研究上的应用进行综述,以期为相关研究提供参考。     

农杆菌介导的花魔芋遗传转化体系的优化

[目的]探讨魔芋愈伤组织培养基、不同转化条件(菌液浓度、侵染时间、预培养时间及共培养时间)及转化植株的筛选条件(抗生素浓度和乙酰丁香酮(AS)浓度)等因素对转化效率的影响。[方法]采用卡那霉素抗性基因为选择标记,对农杆菌介导的花魔芋遗传转化体系进行优化。[结果]在预培养2 d,用OD600为0.6的菌液侵染30 min、共培养3 d,卡那霉素100 mg/L,羧苄青霉素250 mg/L,AS浓度100μmol/L的条件下能有效提高转化效率。再生花魔芋植株经GUS染色及PCR检测,结果表明外源目的基因已经整合到花魔芋基因组中。[结论]为魔芋抗病品种的转基因培育,丰富其种质资源,寻找抗病新途径提供理论依据和试验基础。     

一个简便的适合于分析油菜中SSR位点的检测体系

介绍一个DNA用量少、灵敏度高、重复性好和快速简便的适合于分析油菜中SSR位点的检测体系，该体系对于其它作物的SSR标记分析也有一定的参考价值。     

Development and Characterization of Novel Polymorphic Microsatellite Markers for Tapinoma indicum (Hymenoptera: Formicidae)

Tapinoma indicum (Forel) (Hymenoptera: Formicidae) is a nuisance pest in Asia countries. However, studies on T. indicum are limited, especially in the field of molecular biology, to investigate the species characteristic at the molecular level. This paper aims to provide valuable genetic markers as tools with which to study the T. indicum population. In this study, a total of 143,998 microsatellite markers were developed based on the 2.61 × 10⁶ microsatellites isolated from T. indicum genomic DNA sequences. Fifty selected microsatellite markers were amplified with varying numbers of alleles ranging from 0 to 19. Seven out of fifty microsatellite markers were characterized for polymorphism with the Hardy–Weinberg equilibrium (HWE) and linkage disequilibrium (LD) analysis. All seven microsatellite markers demonstrated a high polymorphic information content (PIC) value ranging from 0.87 to 0.93, with a mean value of 0.90. There is no evidence of scoring errors caused by stutter peaks, no large allele dropout, and no linkage disequilibrium among the seven loci; although loci Ti-Tr04, Ti-Tr09, Ti-Te04, Ti-Te13, and Ti-Pe5 showed signs of null alleles and deviation from the HWE due to excessive homozygosity. In conclusion, a significant amount of microsatellite markers was developed from the data set of next-generation sequencing, and seven of microsatellite markers were validated as informative genetic markers that can be utilized to study the T. indicum population.     

导致牛繁殖障碍的遗传因素及其分析

繁殖障碍的发生几乎都是遗传与环境因素相互作用的结果,因而育种方法有助于减少各种繁殖障碍的蔓延,并且随着牛生产性状遗传进展的累积,必须将在经济重要性方面不低于产肉、产奶等性状的繁殖障碍纳入育种计划中。本文以上述观点为基点,综述了牛的繁殖障碍的遗传性影响因素,介绍了繁殖障碍的几种遗传学分析方法,阐述了控制遗传性繁殖障碍的4项育种措施,提议以寻求适当的标记性状作为繁殖障碍遗传流行病学研究和临床诊断指标的突破口。     

Enhanced bioremediation of soil containing 2,4-dinitrotoluene by a genetically modified Sinorhizobium meliloti

The symbiotic nitrogen-fixing soil bacterium, Sinorhizobium meliloti, is well known for its ability to interact with the leguminous plant Medicago sativa L. It has, however, not been reported that this species possesses the capability to degrade toxic nitroaromatic compounds, such as 2,4-dinitrotoluene (DNT) which is commonly associated with the degradation of the explosive trinitrotoluene (TNT). In this study, the pJS1 DNT-biodegradative plasmid was genetically transferred to S. meliloti strain USDA 1936, which was confirmed by plasmid profile analysis. Several standard analytical and chemical tests including high performance liquid chromatography (HPLC), nitrite (NO₂) release assays, rhizosphere population and plant greenhouse studies were conducted to test the ability of S. meliloti to degrade 2,4-DNT. The possible presence of 2,4-DNT remaining in the treated soil was tested, and no 2,4-DNT had been absorbed by the soil. The pJS1-carrying recombinant strain DHK1 produced ‘ARC’ alfalfa plants that were almost 2-fold higher in shoot dry weight than that produced by the parent strain on soil containing 0.14 mM 2,4-DNT. The transconjugant strain DHK1 reduced significantly one-third more 2,4-DNT in both 0.14 and 0.28 mM contaminated soil, and in 0.55 mM contaminated soil it degraded 94% of the 2,4-DNT present. In liquid cultures, however, only about 4% reduction in 2,4-DNT concentrations was obtained in 10 days. We interpret the results as clearly establishing that genetic modification was successfully used, for the first time, to improve the capability of the symbiotic nitrogen-fixing soil bacterium S. meliloti DHK1 to bioremediate in situ 2,4-DNT-contaminated soil in the presence of alfalfa plants.     

我国甜菜多倍体品种的RAPD分析

利用RAPD分子标记技术对15个甜菜多倍体品种进行了遗传分析。从以前在甜菜中使用过的14个随机引物中筛选出具有非常明显多态性的引物8个,它们在多倍体品种间共扩增出47条不同位置的带,平均5.9条,特异带32条,特异带比率为68%;两个“双引物”共扩增出10条带,公共带仅1条,多态性频率高达90%。15份材料的遗传相似系数在0.75￣0.99。经聚类分析,可将这些材料分成5个组。RAPD聚类结果基本与血缘、系谱及其在生产实践中的表现相吻合,但也有例外。从分子水平上进一步证明了我国糖甜菜育种中存在的遗传材料基础狭窄的问题。