玉米SSR引物在芒属植物遗传多样性分析的应用研究

目的:本文为了探索芒属植物的遗传多样性,方法:运用SSR分子标记的方法,用30对玉米SSR引物对15份中国湖南省的野生芒属材料进行扩增.结果:结果表明:30对引物共扩增出159条带,其中多态性条带121个,占76.10%.平均Nei's基因多样度为0.1992,平均Shannon信息指数为0.3139.UPGMA聚类分析显示,15份材料在遗传相似系数(GS)为0.75水平上,聚成三大类.第一大类由芒组成,第二大类由五节芒组成,第三大类由南获组成.这与形态学上的分类吻合.结论:玉米的SSR引物对于芒属植物遗传多样性分析仍具有很好的可行性.     

Cytological studies on the Sino-Himalayan endemic Anisodus and four related genera from the tribe Hyoscyameae (Solanaceae) and their systematic and evolutionary implications

Cytological studies were carried out on eight species of five genera ( Anisodus , Atropanthe , Hyoscyamus , Mandragora and Przewalskia ) in the tribe Hyoscyameae (Solanaceae). First chromosome counts were reported in six species: Anisodus luridus , A. acutangulus , A. tanguticus , A. carniolicoides , Atropanthe sinensis and Mandragora caulescens , all with 2 n  = 8 x  = 48. Two records, for Hyoscyamus niger (2 n  = 2 x  = 34) and Przewalskia tangutica (2 n  = 4 x  = 44), were also confirmed. All species studied showed the proximal type of mitotic prophase chromosome condensation pattern. Three types of interphase nuclei were recognized: the round prochromosome type for Anisodus , Atropanthe and Mandragora , the rod prochromosome type for Przewalskia and the complex chromocentre type for Hyoscyamus . The cytological data supported the close relationship of all four species of Anisodus . Evidence from the interphase nuclei and chromosome base numbers supported the traditional classification of Hyoscyameae into two groups, i.e. Physochlaina praealta + Hyoscyamus (complex interphase type and x  = 7, 14, 17) and Przewalskia + Atropanthe + Anisodus + Scopolia + Atropa + Mandragora (prochromosome type and x  = 6, 11). Polyploidy is found in most species of the tribe in the Himalayan–Hengduan Mountains, as well as in the Mediterranean region, west–central Asia and eastern Asia. It seems that it probably occurred very early in Hyoscyameae evolution, before the uplift of the Himalayan–Hengduan Mountains. The Himalayan orogeny might have played a minor role in the polyploid evolution of plants in this tribe.  © 2005 The Linnean Society of London. Botanical Journal of the Linnean Society , 2005, 147 , 457–468.     

采用花器喷雾法并基于Cre/lox系统定点整合拟南芥Rps2基因的技术体系研究

To improve site?specific integration technology system, site?specific integration of Rps2 target gene in Arabidopsis thaliana (Linn.) Heynh. was carried out based on Cre/lox system by floral spraying method. The results show that 1495 site?specific integration candidate plants are obtained by this method with a site?specific integration efficiency of about 0076%. After PCR and histochemical staining experiment verification, the positive plants of precise integration account for 8604%, in which, 6334% positive plants are single copy transformed plants. The results of quantitative real?time PCR (qRT?PCR) and hypersensitive reaction (HR) show that the site?specific integrated Rps2 gene can be transcribed and expressed normally. It is suggested that this system can greatly improve the stability and efficiency of site?specific integration genetic transformation system in plants.     

Cytological studies on the Sino-Himalayan endemic Solms-laubachia (Brassicaceae) and two related genera

The karyotypes of six species of Solms-laubachia and one each of Parrya and Desideria (Brassicaceae) from the Himalayan-Hengduan Mountains (Tibet, Sichuan and Yunnan provinces) were investigated. Chromosome counts of S. xerophyta , S. lanata, S. platycarpa and D. baiogoinensis are reported for the first time and found to be diploid with 2 n  = 14. Diploid counts were also made of S. retropilosa , a species previously known to be a tetraploid. The karyotype formulae of D. baiogoinensis , S. eurycarpa , S. pulcherrima , S. retropilosa , S. xerophyta and S. platycarpa are 2 n  = 14 = 6 m + 6sm + 2st, whereas those of S. lanata and P. nudicaulis are 2 n  = 14 = 6 m + 2sm + 6st and 2 n  = 14 = 8 m + 4sm + 2st, respectively. Karyotypes and counts of S. eurycarpa and S. pulcherrima are in agreement with prior studies. The relationships within Solms-laubachia and among the three genera are discussed. Speciation within Solms-laubachia may have been the result of primarily intrachromosomal variation at the diploid level, and the genus probably evolved within the Himalayan-Hengduan Mountains instead of migrating from other regions.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 77−86.     

弱酸（盐）对植物乳杆菌耐酸性的影响

目的探讨弱酸（盐）对植物乳杆菌耐酸性的影响。方法在植物乳杆菌发酵过程中,添加50mmol／L乙酸、10mmol／L丙酸、20mmol／L正丁酸、50mmol／L乙酸钾及2mmol/L柠檬酸钾。结果添加剂量均可使植物乳杆菌耐酸性得到较大的提高,在pH2、37℃下90min,细胞残存率都较对照提高80倍以上,尤以乙酸（盐）、正丁酸为好,而三聚磷酸钾则对细胞耐酸性提高具有较小的作用,细胞残存率较对照约提高5倍。结论在发酵培养基中添加弱酸（乙酸、丙酸及正丁酸）或弱酸盐（乙酸钾、柠檬酸钾及三聚磷酸钾）均可不同程度地提高植物乳扦菌细胞耐酸性。     

温度和渗透应激对植物乳杆菌稳定性的影响

在植物乳杆菌对数期后期,高温（43—47℃）、低温（15—25℃）及10-30g/L NaCl等应激处理60min均可使植物乳杆菌耐热性和耐酸性得到较大的提高。其中,尤以高温应激45℃应激处理效果最好,细胞耐热残存率和耐酸残存率较对照分别提高124％和56．8％。     

山药品种间氨基酸含量的差异性研究

采用盐酸水解法分析了福建省建阳主栽的7个山药品种的氨基酸含量及组成。得出供试材料中都含有17种氨基酸,总氨基酸质量分数在2.86~6.64%,平均4.95%,其中以JY-4品种总氨基酸质量分数最高,JY-2最低;各种氨基酸质量分数高低顺序基本相似,以精氨酸、谷氨酸、天冬氨酸居前三位,胱氨酸质量分数最低,必需氨基酸质量分数的高低依次是亮氨酸>苯丙氨酸>赖氨酸>缬氨酸>异亮氨酸>苏氨酸>蛋氨酸;鲜味氨基酸在各品种间的差异显著,以JY-4最高,达3.41%,依次是JY-3、JY-6、JY-1、JY-5、JY-7、JY-2。再运用聚类分析方法从氨基酸的组成及含量角度对7个品种进行分类可分为三类;相关分析表明:山药总氨基酸含量愈高,必需氨基酸和鲜味氨基酸含量亦愈高。     

双向标记培养植物测定大气二氧化碳稳定碳同位素组成

基于植物能够利用体内的碳酸酐酶来催化碳酸氢根离子生成二氧化碳和水作为底物进行光合作用的特性,采用两种δ13CPDB值差值大于10‰的碳酸氢钠分别作为外源碳酸氢根离子的碳同位素标记物,通过室内双向水培诸葛菜和芥菜型油菜实验,分别向水培处理液里添加已知δ13CPDB值的碳酸氢钠并培养24 h,利用同位素比值质谱(IRMS)技术,测定并计算了两个时间、两种环境下的大气二氧化碳稳定碳同位素日平均组成。结果表明:在环境1(不同浓度的Na HCO3处理液)下所得到的δCa值与添加到处理液中的碳酸氢根离子的浓度有关;在环境2(不同浓度的PEG处理液)下所得到的δCa值与添加到处理液中的PEG的浓度无关;两种环境下所测得的大气二氧化碳稳定碳同位素日平均组成δCa值与实验中培养的植物种类无关,而与添加到培养液中碳酸氢根离子的浓度及植物的生长速率有关。数据重现性好,结果准确可靠,可以高精度的测定不同待测环境下大气二氧化碳稳定碳同位素比值,其可为以后监测不同时间、不同地点的大气二氧化碳碳同位素组成及来源提供非常有效的方法和信息。     

猪繁殖与呼吸综合征病毒GP5a多克隆抗血清的制备

构建XX2012株猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)GP5a蛋白基因的真核表达载体,并制备出GP5a蛋白的多克隆抗血清。以XX2012株PRRSV GP5a蛋白的基因序列为扩增模板,设计并合成一对特异性扩增引物,通过RTPCR扩增得到该病毒株的GP5a蛋白全长基因片段,将其定向克隆到真核表达载体p CAGG中,构建含有GP5a蛋白基因的重组表达载体p CAGGS-GP5a,并将获得的p CAGGS-GP5a载体采用基因免疫的方式免疫BALB/c小鼠制备GP5a蛋白的多克隆抗体。结果显示,成功克隆出了XX2012株PRRSV GP5a蛋白全长基因,片段大小为156 bp;构建的p CAGGS-GP5a载体经PCR、双酶切、测序鉴定均无误;通过三次基因免疫小鼠成功制备出了GP5a蛋白的多克隆抗体,间接免疫荧光试验证实制备出的多克隆抗体能特异性识别病毒感染后的细胞。由此获得了PRRSV GP5a蛋白基因的真核表达载体,制备出了GP5a蛋白的特异性多克隆抗血清,为今后开展GP5a蛋白的亚细胞定位及相关功能的研究奠定了基础。     

薯蓣属植物人工杂交后代的检测

以湖北武当山的盾叶薯蓣(Dioscorea zingiberensis C.H.Wright)为主要亲本,与重庆金佛山的盾叶薯蓣、小花盾叶薯蓣(D.parviflora C.T.Ting)及黄独(D.bulbifera L.)进行种内和种间杂交,并获得部分杂交组合的F1代植株。将F0代种子与F1代植株进行结实率及种子萌发率比较,并与亲本进行过氧化物酶(POD)、超氧化物歧化酶(SOD)和酯酶(EST)同工酶比较。结果表明,不同亲本及其杂交后代的结实率、种子萌发率和实生苗存活率均存在明显差异;亲本和F1代的同工酶酶谱较丰富,部分杂种与亲本的相似度较高,并出现各自的特征酶带。运用同工酶技术可以鉴别杂种的真伪。