基于DNA条形码和SRAP的太子参种质遗传多样性分析

应用DNA条形码和SRAP分子标记技术,开展15份太子参种质资源遗传差异分析。太子参DNA条形码显示：ITS1序列的突变位点占比最多为2.16%,rbcL6序列的突变位点占比最少为1.23%;psbA-trnH序列缺失位点占比最多为4.88%;ITS1序列的GC含量最高,达55.52%;psbA-trnH序列的GC含量最低,仅为24.71%。太子参SRAP标记筛选出9对引物,扩增出215个位点,其中42个为多态性位点,多态性位点百分率（PPL）为19.53%;从扩增位点总数看,多态性条带丰富、清晰,既有共同位点又有特异性位点;7个居群Nei’s遗传多样性指数（H）范围为0.3466～0.4985,Shannon信息指数（I）范围为0.5301～0.6917,显示出较高的遗传多样性水平,太子参种群基因多样性（Ht）为0.4004,其中种群内基因多样性（Hs）和遗传分化系数（Gst）分别为0.0901、0.3103,分别占Ht的77.50%、22.50%。种源间Gst为0.7506,即有75.06%的遗传变异存在于种源间,24.94%的遗传变异存在于种源内,表明太子参种源间遗传变异大于种源...     

基于EST-SSR毛细管电泳荧光标记技术分析广东2个历史名茶群体遗传多样性

利用EST-SSR毛细管电泳荧光标记技术对罗坑和马图2个广东历史名茶茶树群体的共52份种质资源进行研究。结果表明:27对引物共检测到204个位点,平均每对引物检测7.56个位点,引物检测到的基因型数为11.22个;观测等位基因数为7.56,有效等位基因数为3.42;观测杂合度和期望杂合度平均值分别为0.55和0.66;多态性信息指数(PIC)平均值为0.61;反应遗传多样性的Shannon’s遗传信息指数为1.40。2个茶树群体的基因流Nm平均值为3.19,表明2个群体间在过去的某个时间可能有过基因交流,群体间遗传分化系数(Fst)的平均值为0.072 7;2个亲体的遗传多样性水平相近。依据Evanno统计模型分析可将52份茶树种质分为3个亚群,分别包含18、24、9份茶树种质资源,MT4种质没有明确的亚类特性。      

广西甘蔗花叶病SCMV调查初报

为探明甘蔗花叶病SCMV在广西蔗区的发生情况,给甘蔗健康种苗生产提供科学依据,从广西各主要蔗区对不同品种采集表现花叶症状的甘蔗样品,采用1对病毒CP基因引物(SCMVF4:5′-GTTTTYCACCAAGCTGGAACAGTC-3′;Y=CorT,SCMVR3:5′-AGCTGTGTGTCTCTCTGTATTCTC-3′),进行一步法RT-PCR检测。结果表明:69.4%的样品为阳性。选取8份代表性样品,对经SCMVF4/SCMVR3扩增获得的病毒CP基因片段进行直接测序,所得序列经Blast比对确认均为SCMV CP序列。在所调查的品种中,主栽品种ROC22最易受SCMV侵染,其它台糖系列如台糖16、台糖28、台糖95-889、台优,柳城03-182、柳城03-1137及广西甘蔗研究所许多品系均检测到SCMV。目前甘蔗花叶病SCMV已在广西主要蔗区普遍发生。     

基于nSSR和cpDNA序列的城步峒茶群体遗传多样性和结构研究

采用简单重复序列标记(nSSR)与叶绿体DNA序列(cpDNA)分析技术,对城步峒茶群体进行了遗传多样性、遗传结构和遗传分化等研究。结果表明,15对nSSR引物在参试81份资源中共获得142个等位位点,平均每对引物9.47个,城步峒茶群体的观测杂合度(Ho)、期望杂合度(He)和Nei期望杂合度(Nei)分别为0.49、0.62和0.62,具有较高的遗传多样性。Structure分析将79份峒茶资源分成3个类群,但各类群的遗传背景较为复杂,没有明显的群体结构。F检验表明,城步峒茶群体的近交系数F_IS为正值(F_IS=0.177 5),群体间的遗传分化系数F_ST较小(F_ST=0.034 5),分化程度较低,基因流Nm较高(Nm=7.01)。3对cpDNA引物分别获得了473 bp(rbcL)、704 bp(matK)和320 bp(psbH-trnA)的片段序列,变异位点占总位点的比例分别为0.42%、0.71%和1.25%。将3个序列依次拼接,共产生了9个单倍型,单倍型数由多到少的居群依次为TXZ(6)、DZC(4)、DPS(4)、TYS(3)、HJZ(2),群体的单倍型多样性(Hd)和核苷酸多样性(π)分别为0.732和0.001 39。9个单倍型中,单倍型H1和H5处于进化网络图的中心节点上,并且包含资源数量最多,属于比较原始的单倍型。同时,nSSR和cpDNA的AMOVA分析结果基本一致,居群内的变异百分比分别达到96.69%和80.54%,城步峒茶的遗传变异主要存在于居群内。     

应用RT-PCR检测水仙潜隐病毒

根据已报道的水仙潜隐病毒(Narcissus latent virus,NLV)外壳蛋白(Coat protein,CP)基因序列,设计一对特异性引物,以提取的水仙样品总RNA为模板进行RT-PCR,并进行特异性和灵敏度测定。结果表明,此法能够从携带NLV的水仙样品中成功扩增出大小约829 bp的特异性目的片段,具有良好的特异性和较高的灵敏度,可有效地应用于水仙样品中NLV的检测。     

云南新平蔗区发现甘蔗白条病

利用白条黄单胞菌[Xanthomonas albilineans (Ashby) Dowson]特异性引物XAF1/XAR1对2019年采自云南新平蔗区的8份疑似甘蔗白条病样品进行PCR分子检测,结果表明所有样品均扩增得到大小约608 bp的特异性条带,GenBank登录号为MT625864～MT625871。这些核苷酸序列与白条黄单胞菌(X. albilineans)GPE PC73菌株基因组上的三磷酸腺苷结合盒(Adenosine triphosphate-biding cassette,ABC)转运蛋白基因abc(基因号XALc_1791)的序列一致性为100%。系统发育树分析表明这些白条黄单胞菌菌株序列处于同一进化分支。结合田间症状诊断和分子鉴定结果,确认云南新平蔗区发生的甘蔗病害为X. albilineans引起的甘蔗白条病,这是云南新平蔗区首次发现。     

吉林省不同产地和采收时间对白屈菜有效成分含量的影响

目的通过对比吉林省不同产地及采收时间的白屈菜有效成分含量,研究采收时间和产地对白屈菜有效成分含量的影响。方法采用高效液相色谱法(High Performance Liquid Chromatography,HPLC)检测吉林省不同产地和采收时间白屈菜中白屈菜红碱的含量。结果不同产地、采收时间白屈菜样品之间的白屈菜红碱含量均有差异,在54个白屈菜样品中,有28个样品的白屈菜红碱含量低于0.02%,不符合2015年版《中国药典》标准。结论采收时间和产地均会影响吉林省范围内白屈菜的有效成分含量,其中7月份靖宇地区采集的白屈菜中有效成分最高。     

进口大豆中菜豆晕疫病菌巢式PCR检测方法的建立

为实现对进口大豆样品中菜豆晕疫病菌的快速检测,本研究根据菜豆晕疫病菌的argK特异性序列设计引物52B/8F和24B/24F,建立了巢式PCR检测方法,并进行特异性和灵敏度验证。试验结果表明,利用此检测方法对多种参试菌株进行检测时,只有菜豆晕疫病菌呈阳性反应,而其他病菌不产生扩增反应;巢式PCR检测方法对菜豆晕疫病菌基因组DNA和菌悬液检测时,其灵敏度分别达到0.916×10-4ng/μL和2.4×103CFU/m L,比常规PCR灵敏度高1 000倍。在对100份进口大豆样品检测时,3份样品扩增到了特异性条带,测序分析结果证明3份大豆样品中携带的病菌确实为菜豆晕疫病菌。本研究所建立的巢式PCR检测方法具有特异性强、灵敏度高、检测时间短并且检测准确率高等特点,可用于口岸菜豆晕疫病菌的检测。     

基于SLAF-seq技术的红心火龙果SNP位点开发及遗传分析

为分析不同品种红心火龙果的遗传关系,本研究采集国内14种红心火龙果品种,通过生物信息学分析进行实验方案的系统设计,将所提取基因组进行酶切,筛选特异长度的DNA片断,构建SLAF-seq文库后进行高通量测序,将获得的序列进行分析比较后得到多态SLAF标签,在此标签基础上进一步开发特异性SNP位点,最后建立14种红心火龙果品种的进化树。本研究共开发159 560个SLAF标签,样品的平均测序深度为5.26×;共得到120 294个群体SNP,其中高一致性的群体SNP 51 859个。12种红心火龙果聚类结果发现,样品1、2、6、8、10同缘关系相近;样品3、5、7、9同缘关系相近;样品4、14同缘关系相对较近。研究表明应用SLAF-seq技术开发分子标记的效率比ISSR、RAPD、AFLP等常规技术更高。12种红心火龙果的聚类结果从基因组水平揭示不同地区品种之间的遗传分化关系,为火龙果种质的保护和遗传改良提供参考。     

NR/ENR/SiO_2复合材料的阻尼性能研究

采用机械混炼的方法制备不同ENR含量的NR/ENR/SiO2复合材料,使用动态热机械分析仪(DMA)和橡胶加工分析仪(RPA)对NR/ENR/SiO2复合材料的动态力学性能和阻尼性能进行分析。结果表明:ENR对NR/ENR/SiO2复合材料的阻尼性能具有显著影响。动态热机械分析测试结果表明,随着ENR用量的增多,NR/ENR/SiO2复合材料在使用温度范围内的损耗因子积分面积增大,阻尼性能提高。橡胶加工分析频率扫描和应变扫描测试结果表明,在频率小于10 HZ不同应变时,加入了ENR的NR/ENR/SiO2复合材料的阻尼因子均高于NR/SiO2复合材料。扫描电镜分析结果证实,ENR的加入减少了SiO2的自聚,改善了填料在橡胶基体中的分散,从而使NR/ENR/SiO2复合材料力学性能得到提高。     

纳米TiO_2/天然橡胶复合材料的抗菌性研究

采用溶胶凝胶法,以钛酸四丁酯掺杂金属Ag制备纳米二氧化钛(Ag-TiO2)水溶胶,并与天然胶乳湿法共混制备得到纳米TiO2/天然橡胶复合材料。紫外可见光谱表明,金属Ag能提高TiO2的光催化性能;透射电子显微镜观察到TiO2颗粒粒径为50nm左右,并且均匀的吸附在胶粒表面。研究此复合材料的抗菌性能结果表明,其具有良好的抗细菌和抗霉菌效果,对大肠杆菌的抗菌率达到90%以上,特别是硫化过后的纳米TiO2/天然橡胶复合材料的抗细菌率更达98.5%。     

In VitroDigestion of Wheat Microstructure with Xylanase and Cellulase fromTrichoderma reesei

Resistant starch (RS), producedin vitroby hydrolysis of retrograded pea starch gels and amylose gels by porcine pancreaticalpha-amylase, was characterised by X-ray diffraction, size exclusion chromatography and methylation analysis. These techniques showed that RSin vitroconsisted of semi-crystalline, mostly linear material that was present in two main molecular size subfractions (DP_n>100 andDP_n20–30) with a third, minor subfraction (DP_n≤5). The extent of retrogradation of amylose was found to be of primary importance in determining the RS content of starch. Analysis ofin vivoRS, recovered during an ileostomy study, produced results that were similar to those obtained from RSin vitro. Anin vitromodel for the structure of resistant starch is proposed.     

Physicochemical Studies on Resistant StarchIn VitroandIn Vivo

Resistant starch (RS), producedin vitroby hydrolysis of retrograded pea starch gels and amylose gels by porcine pancreaticalpha-amylase, was characterised by X-ray diffraction, size exclusion chromatography and methylation analysis. These techniques showed that RSin vitroconsisted of semi-crystalline, mostly linear material that was present in two main molecular size subfractions (DP_n>100 andDP_n20–30) with a third, minor subfraction (DP_n≤5). The extent of retrogradation of amylose was found to be of primary importance in determining the RS content of starch. Analysis ofin vivoRS, recovered during an ileostomy study, produced results that were similar to those obtained from RSin vitro. Anin vitromodel for the structure of resistant starch is proposed.     

细旦涤麻棉针织物的纤维素酶整理

本文介绍了酶整理技术在细旦涤麻棉针织物上的应用，提出了现在主要三种纤维素酶处理细旦涤麻棉针织物的最佳工艺及效果。     

巴西橡胶树AnnHb1基因的克隆及表达分析

根据一个从巴西橡树胶乳cDNA文库中获得的EST片段的序列设计引物,通过RACE的方法获得了橡胶树编码annexin的cDNA,命名为AnnHb1。序列分析表明,AnnHb1长为1 198 bp,含有945 bp的阅读框,62 bp的5'-UTR和191 bp的3'-UTR,编码314个氨基酸,分子量为35.99 ku,等电点为8.18。该氨基酸序列与蓖麻、麻疯树、棉花、苜蓿、烟草和拟南芥中的annexin的同源性分别为82%、72%、72%、64%、60%和60%。半定量RT-PCR分析结果表明AnnHb1基因在愈伤、花、树皮、叶、胶乳中均有表达,其中在愈伤组织中表达量最低,树皮中表达量最高。     

Polymorphism of Low Mr Glutenin Subunits in Triticum tauschii

A collection of 173 Triticum tauschii accessions was analysed to evaluate the variability of low molecular weight (M_r) glutenin subunits. These proteins were analysed by one-step one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and were divided into B-, C- and D-subunits in accordance with their electrophoretic mobility. Extensive polymorphism, both in the number and electrophoretic mobility, was detected in lowM_r glutenin subunits present in T. tauschii. Thirty different patterns for B-subunits and forty-three for C-subunits were identified, some of which were with identical electrophoretic mobility than those observed in hexaploid wheat. Glutenin subunits with the same electrophoretic mobilities of low M_r D-glutenin subunits as well as subunits encoded at the Glu-D4 and Glu-D5 loci, were also detected in accessions of T. tauschii. These results provide new basic knowledge regarding the genetics variability of the low M_r glutenin subunits, as well as their potential to create novel germplasm for the improvement of wheat quality in breeding programs.     

Comparative Enzyme Kinetics of Two Allelic Forms of Barley (Hordeum vulgare L.) Beta -amylase

The barley (Hordeum vulgare L.) varieties, Franklin and Schooner, contain two different allelic forms of beta -amylase (EC 3.2.1.2) encoded on chromosome 4H by the Bmy 1-Sd1 and Bmy 1-Sd2L alleles, respectively. The corresponding enzymes, referred to as Sd1 and Sd2L, were purified from both mature barley grain and germinated barley (green malt), and their physical and kinetic properties studied. Approximately 4 kDa were cleaved from both Sd1 and Sd2Lbeta -amylases after germination. The K_mvalue for green malt beta -amylase was less than that of mature grain beta -amylase for both varieties when potato starch was used as a substrate, although V_maxwas similar. This indicated that proteolysis after germination increased the affinity of beta -amylase for potato starch. No significant kinetic differences were observed between beta -amylase from mature grain and green malt of the two barley varieties when amylose (degree of polymerisation 100 and 18) and maltopentaose were used as substrates. Kinetic differences were also observed between the two allelic forms of beta -amylase. Sd1 beta -amylase from green malt exhibited a lower K_mvalue for potato starch than Sd2L beta -amylase, demonstrating that at non-saturating starch concentrations Sd1 beta -amylase is better able to hydrolyse starch than Sd2L beta -amylase. As the degree of polymerisation of the substrates decreased from approximately 740 (potato starch) to 5 (maltopentaose), the K_mvalues for beta -amylase increased, whereas V_maxvalues decreased. Maltose, the hydrolytic product of beta -amylase, was found to be a weak competitive inhibitor of both Sd1 and Sd2L green malt beta -amylases with respect to potato starch and amylose. Taken together the kinetic observations for bet a-amylase suggest that the allelic differences and C-terminal proteolysis might be exploited to improve the efficiency of starch hydrolysis during the mashing stage of the brewing process.     

杜鹃红山茶离体快速繁殖

以杜鹃红山茶实生小苗为试验材料,探讨离体快速繁殖的方法。结果表明:茎段外植体在MS+BA 1 mg/L+IBA 0.01 mg/L+GA3 10 mg/L的培养基上培养60 d,侧芽萌发率高达68%;将这些萌发的侧芽连同原来的茎段外植体移至添加BA 0.2～0.4 mg/L的培养基培养1个月,74%的侧芽能够继续生长并形成丛生芽;再将丛生芽移至1/2 MS+IBA 4 mg/L+0.1%活性炭的培养基上培养1个月,得到大量伸长的芽条;将芽条切离母体后插植于1/2 MS+IBA 8 mg/L的培养基,60 d后生根率达到90%。通过以上过程繁殖得到的小苗质量良好,移栽1个月后的成活率可达90%。     

Protein Inhibitors ofAlpha-amylase in Mature and Germinating Grain of Rye (Secale cereale)

The activities of endogenous (R-type) and exogenous acting (D-type) protein inhibitors ofalpha-amylase and the activities ofalpha- and total amylase were determined in milling fractions of rye. High D-type amylase inhibitor activities were detected in the embryo (255 IU/g) and in the endosperm fraction (64·9 IU/g), low inhibitor activities were found in the aleurone layer fraction (25·9 IU/g). The highest R-typealpha-amylase inhibitor activity was found in the aleurone layer fraction (32·6 IU/g), and the lowest value in the epidermis containing fraction (5·0 IU/g). The D- and R-typealpha-amylase inhibitor activities varied with growing conditions. D-type amylase inhibitor activities were found to be high in those samples which grew under drought conditions and low in samples cultivated under wet and cool weather. Higher R-typealpha-amylase inhibitor activities were found in rye genotypes cultivated under wet conditions and lower values under dry weather. There were small variations inalpha-amylase inhibitor activities between sprout-stable and sprout-sensitive rye genotypes. The D- and R-typealpha-amylase inhibitor activities of all varieties were stable during 72 h of germination. Similar soil conditions will therefore lead to differentialalpha-amylase inhibitor activities depending on weather conditions during growth.