23株豆科木本植物根瘤菌16SrDNA序列分析

为研究和利用华东地区豆科植物根瘤菌的种质资源多样性,对从华东地区豆科木本植物根瘤分离获得的23个菌株进行了16S rDNA全序列分析,并与相关参比菌株的16S rDNA序列进行了比较及聚类分析,建立了23株华东地区豆科木本植物根瘤菌的发育树状图,这一分析结果与来自形态学的鉴别结果吻合,初步确定了23株豆科木本植物根瘤菌分类地位.     

两株花生根瘤菌的比较研究及结瘤能力的测定

从花生根瘤中筛选出两株生长速度差异明显的花生根瘤菌:1K(快生型),2M(慢生型),通过16S rDNA测序分析并结合生理生化实验确定两种根瘤菌分别为中华根瘤菌和慢生根瘤菌。对二者的生理特性、抗逆性和结瘤能力等指标进行了研究,发现慢生型虽然在生长速度、抗逆性等方面明显不及快生型,但是结瘤能力比快生型要强。     

土壤中一株产黑色素微生物的分离与鉴定

以土壤为材料,分离纯化得到了一株产黑色素的微生物(D-1),根据菌株的形态特征并结合16S rDNA序列比对分析,确定这株产黑色素的微生物属于链霉菌属(Streptomyces).并测定了菌株的生长曲线、抗生素抗性、所产黑色素的吸收光谱,为后期优化产黑色素的条件提供了理论基础.     

嗜热菌分离筛选及分子分类初探

从厦门周边地区温泉采集得到样品,在75℃和80℃的温度下进行培养并分离得到60株嗜热菌.为了从大量的菌株中筛选出不同的菌株,有效地简化筛选过程,应用了细菌全蛋白SDS-PAGE电泳、RAPD和16S-23S rDNA间隔区比较分析等分子生物学方法从中筛选得到7株种属不同的嗜热菌,并进一步运用16S rDNA序列分析鉴定其种属关系,为深入研究奠定了基础.     

一株无机磷细菌的分离、鉴定及其溶磷能力分析

从根系土壤分离纯化出降解无机磷的细菌8株,通过平板法进行初筛,根据水解圈直径和菌落直径比值大小筛选到水解圈直径和菌落直径比值较大的菌株3株,进一步通过液体摇瓶培养复筛出I株分解无机磷能力最强的菌株PC-2,该菌株经形态特征,生理生化特性16 S rDNA序列测定等一系列试验鉴定为地衣芽孢杆菌(Bacillus lich...     

一株拮抗姜瘟青枯劳尔氏菌的泛菌的分离及鉴定

从生姜田土中分离到一株对姜瘟青枯劳尔氏菌(Ralstonia solanacarum)有强拮抗作用的泛菌菌株Q6，对该菌株进行了形态特征、培养特征、生理生化特征的鉴定及16S rDNA 序列的分析.以16S rDNA序列为基础构建了包括12 株相关种属细菌在内的系统发育树，其中与11个泛菌属的菌株的16S rDNA序列的同源性为95.54%～99%.     

一株广谱抗菌活性菌株的鉴定及生物学特性研究

从湛江湖光岩世界地质公园的火山岩石中分离到一株具有广谱抗菌活性的菌株,命名为菌株HGY25,该菌株对多种食品致病菌均有较强的抑制作用。通过形态学观察、生理生化指标测定、16S rDNA碱基序列测定和同源性分析,鉴定该菌株为枯草芽孢杆菌Bacillus subtilis。对菌株部分生物学特性研究表明:其生长的最适pH值为7.0,最适生长温度为37℃。     

有机磷降解菌的分离筛选及初步鉴定

目的:从不同根系土壤分离筛选高效降解有机磷的细菌.方法:通过平板法进行初筛,根据水解圈直径和菌落直径比值大小筛选到水解圈直径和菌落直径比值较大的菌株,进一步通过液体摇瓶培养复筛,对其解磷能力进行了测定,并通过形态特征、生理生化特性和16 S rDNA序列测定等一系列试验时其进行鉴定.结果:分离纯化出10株解磷菌,经平板...     

具有AHL降解能力的海洋微生物的筛选与鉴定

以紫色视杆菌Chromobacterium violaceum 026为报告菌株,以己酰基高丝氨酸内酯作为菌株生长的唯一碳源及能源,通过富集培养、分离纯化,从西南印度洋海泥中筛选得到2株菌S2和S3。检测结果表明,S2、S3可能具有胞内AHL降解酶活性。形态鉴定和16S rDNA序列分析表明,2株菌均为芽孢杆菌属。系统发育分析表明,菌株S2、S3与Bacillus amyloliquefaciens的亲缘关系最近。     

Nitro苜蓿和鹰嘴紫云英根瘤菌的分离与鉴定

采用划线和稀释分离相结合的方法对美国Ｎｉｔｒｏ苜蓿和鹰嘴紫云英根瘤的分离物进行纯化，并对分离的菌株进行了鉴定及理化测试，研究结果表明，这两种根瘤菌株生长速度快，结瘤率高，具有较强的耐盐性，其耐受浓度达到０．６ｍｏｌ／Ｌ和０．７ｍｏｌ／ＬＮａＣｌ，从鹰嘴紫云英的根瘤中分离的根瘤菌株在ＢＴＢ培养基中培养产酸，在石蕊牛奶培养基中培养产碱。     

野大豆与栽培大豆愈伤组织耐盐性比较

野大豆(Glycine soja)是唯一能与栽培大豆(Glycine max)杂交而后代能育的野生近缘种,具有极强的耐盐碱性,利用野大豆与栽培大豆有性杂交,结合离体组织培养技术,进行耐盐性变异体的离体筛选,是提高大豆耐盐性的一种有效途径。本研究选择5种栽培大豆品种和从黄河三角洲地区(山东东营)获得的野大豆,进行了大豆成熟胚愈伤组织诱导及愈伤组织耐盐性研究。结果显示,不同浓度Na Cl胁迫下,野大豆愈伤组织的存活率明显高于栽培大豆,与对照相比鲜重下降幅度明显低于栽培大豆;Na Cl胁迫下,野大豆愈伤组织的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性明显高于栽培大豆愈伤组织,MDA含量明显低于栽培大豆,表明野大豆愈伤组织耐盐性明显优于栽培大豆。该研究为耐盐大豆新品种培育提供了理论依据。     

金华地区野生大豆小种群的分子生态学研究

为了阐明小地区范围内野生大豆种群的遗传分化情况,应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）方法,对浙江金华地区５个一年生野生大豆种群,进行了分子生态学研究。用根据ＲＡＰＤ数据计算得 Ｓｈａｎｎｏｎ指数估计了５个野生大豆种群的遗传多样性。发现大部分的遗传变异存在于野生大豆种群间,只有少部分的遗传变异存在于种群内。     

不同纬度野生大豆种群间的遗传变异

选用了经初步筛选的２０个短随机序列引物和２个长随机序列引物对５个不同纯度来源的野生大豆进行了ＤＮＡ随机扩增,其中有１９个引物扩增出多态性片段,区获得７０个ＲＡＰＤ多态性片段,平均每个引物３．６８条,运用自编的计算机程序对扩增进行聚类分析。     

Genetic diversity, geographic differentiation and evolutionary relationship among ecotypes of Glycine max and G. soja in China

The knowledge of origin and evolution of cultivated soybeans is one of the basic issues in both biology and agronomy of the crop. In order to investigate the nuclear and cytoplasmic genetic diversity, geographic differentiation and genetic relationship among geographic ecotypes of cultivated （Glycine max） and wild （G. soja） soybeans, the allelic profiles at 60 nuclear simple-sequence repeat （nuSSR） loci and 11 chloroplastic SSR （cpSSR） loci evenly distributed on whole genome of 393 landraces and 196 wild accessions from nation-wide growing areas in China were analyzed. （i） The genetic diversity of the wild soybean was obviously larger than that of the cultivated soybean, with their nuSSR and cpSSR alleles as 1067 vs. 980 and 57 vs 44, respectively. Of the 980 nuclear alleles detected in the cultivated soybean, 377 new ones （38.5%） emerged, while of the 44 chloroplastic alleles in the cultivated soybean, seven new ones （15.9%） emerged after domestication. （ii） Among the cultivated geographic ecotypes, those from southern China, including South-Central China, Southwest China and South China possessed relatively great genetic diversity than those from northern China, while among the wild geographic ecotypes, the Middle and Lower Changjiang Valleys wild ecotype showed the highest genetic diversity. （iii） The analysis of molecular variance, association analysis between geographic grouping and molecular marker clustering and analysis of specific-present alleles of ecotypes demonstrated that the geographic differentiation of both cultivated and wild soybeans associated with their genetic differentiation, or in other words, had their relevant genetic bases. （iv） The cluster analysis of all accessions clearly showed that the wild accessions from Middle and Lower Changjiang Valleys and South-Central ＆ Southwest China had relatively small genetic distances with all cultivated accessions. The UPGMA dendrogram among geographic ecotypes further showed that the genetic distances between all cultivated ecotypes and the Middle and Lower Changjiang Valleys wild ecotype were smaller than those with other wild ones, including their local wild counterparts. Therefore, it is inferred that the wild ancestors in southern China, especially those from Middle and Lower Changjiang Valleys might be the common ancestor of all the cultivated soybeans.     

Isolation and application of effective nitrogen fixation rhizobial strains on low-phosphorus acid soils in South China

Soybean (Glycine max L.) is a very important food and oil crop in China. Legume-rhizobium symbiotic nitrogen (N) fixation is an important biological character and also the base of improving soil fertility of soybean. However, soybean production and development is severely limited in tropical and subtropical areas in China due to a lack of effective rhizobial inoculants adapting to low-phosphorus (P) acid soils. In the present study, 12 soybean rhizobial strains were isolated and purified from the nodules of two soybean genotypes contrasting in P efficiency, which were grown on different low-P acid soils with different soybean cultivation histories. Results from 16S rDNA sequence analysis showed that these 12 rhizobial strains belonged to the genus of Bradyrhizobium, which had higher nitrogenase activities compared to the control strain, Bradyrhizboium japonicum USDA110. A field experiment was carried out by applying rhizobial inoculants, a mixture of three rhizobial strains that showed the highest nitrogenase activity, on a typical low-P acid soil in South China. The results showed that, without inoculation, no nodules were formed in the three soybean genotypes tested; with inoculation, the nodulation rates in all were 100%. Inoculation with rhizobial inoculants not only made many nodules formed, but also increased soybean shoot biomass and yield, and improved nitrogen (N) and P nutrient status. Among which, shoot dry weight, N and P content of a soybean genotype, Huachun 3, inoculated with rhizobium were increased 154.3%, 152.4% and 163.2% compared to that without inoculation, respectively. We concluded that: (i) The effective indigenous rhizobial strains isolated in this study from soybeans on low-P acid soils in South China have the characters of broad host range, high nodulation efficiency, efficient N fixation, great low pH and low P tolerance. (ii) Soil environment and host types are the key factors to screen the effective rhizobial strains. Considering soil pH values and P efficiency of the host genotypes might increase the screening efficiency. (iii) Improving N status and facilitating root growth might be the mechanisms of increasing the P uptake in soybean plants inoculated with the effective rhizobial strains on low-P acid soils. (iv) Inoculation with the effective rhizobial inoculants could significantly improve growth, N and P content of soybean on low-P acid soils, which might be an effective approach to enhance soybean cultivation and development in these areas. Therefore, application and extension of inoculation techniques with effective rhizobial inoculants in legumes would result in great economical, environmental and ecological benefits. The authors contributed equally to this work Supported by National Key Basic Research and Development of China (Grant No. 2005CB120902), McKnight Foundation Collaborative Crop Research Program (USA) (Grant No. 05-780) and National Natural Science Foundation of China (Grant No. 30571111)     

铝对大豆一些生长特性的影响

以2个大豆(Glycine maxMerrill.)品种华春18、浙春3号为材料,设置不同浓度的铝处理进行土培,研究铝对大豆主要生长特性的影响.结果表明:适量的铝处理对大豆生长具有一定的促进作用;高浓度铝会对大豆产生不利影响,阻碍大豆株高、茎粗、叶面积、比叶重的增长.2个品种比较,浙春3号抗铝胁迫的能力大于华春18.     

野生大豆DNA导入小麦及RAPD分子验证

利用花粉管通道法将野生大豆总DNA导入小麦，以期获得变异系小麦．对小麦球蛋白的SDS—PAGE，印迹表明，冬小麦T2l6#产生了新的蛋白亚基，其分子量为78kD，而有的变异品系一些蛋白亚基消失了；凯氏定氮法和氨基酸分析表明小麦后代T216#的总蛋白含量增加，其氨基酸组成，尤其是赖氨酸含量明显提高．RAPD分析结果表明，新品系基因组出现多态性，并具有供体特异性DNA带，这些变异现象表明该小麦品系为转基因后代．本实验为选育高蛋白、优质的新小麦品种提供了途径．     

Genetic variations of glycinin subunit genes among cultivated and wild type soybean species

Glycinin is a predominant storage protein in most soybean accessions. It is a hexamer constituted by five major subunits, which can be classified into two groups. Group I contains G1, G2 and G3, and Group II contains G4 and GS. The genes encoding these subunits have been designated from Gyl to Gy5, respectively. In the present study, Gyl genomic fragments were cloned from wild accessions of subgenera Glycine glycine, Glycine soja and a cultivar of Glycine max. Their sequences and the deduced amino acid sequences were compared. The residues critical for assembling of G1 subunits from the wild perennial accession were conservative. The Gy4 fragments were cloned from two wild perennial accessions and compared with that from subgenus Soja. The intron 3 of Gy4 had abundant variations between the subgenera G. soja and G. glycine as well as within the subgenus G. glycine. Abundant variations existed in the disordered regions 3 and 4 of G4 subunits from two wild perennial accessions. The genomic organization of glycinin genes was analyzed in 19 accessions from subgenera Soja and Glycine. The hybridization patterns were identical among the accessions of subgenus Soja. On the contrary, abundant polymorphisms existed between the accessions from subgenus Glycine. These results indicated that glycinin genes have high degree of conservation within subgenus Soja but more variations within subgenus Glycine.     

soja亚属的数量分类学研究

以不同进化类型的7 个大豆居群为分类运算单位,选取了28 个形态学、生态学性状,对不同进化类型的大豆居群进行了数值分类研究.Q 分析及图论分析揭示野大豆、半野生大豆与栽培大豆之间存在较大差异.主成份分析表明前三主成份可保留总信息量的91 .05 % ,说明种内居群分类所选性状合理,并能在三维空间表达各居群间的相对位置.