离子注入法导入外源DNA引起陆地棉性状变异的研究

通过３０ｋｅＶ不同剂量的Ａｒ＾＋注入和ＤＮＡ溶液浸滴法将比克氏棉和红麻ＤＮＡ导入泗棉２号。结果表明：离子注入可显地促进外源ＤＮＡ导入受体，增加受体后代性状的变异频率和变异类型，而且多数变异可迅速趋于稳定。在供试剂量中，以２×１０＾１５Ａｒ＾＋／ｃｍ＾２介导外源ＤＮＡ引起受体性状变异的频率最高，达１６．２％。经选育已获得了一批抗枯萎病、早熟、种子少腺体和优质等各具特色的陆地棉新种质系。     

定量测定甲胺磷残留的间接竞争ELISA的建立和初步应用

以水溶性碳化二亚胺法将甲胺磷（ＭＴＰ）分子偶联牛血清白蛋白（ＢＳＡ）上,用此合成的免疫原复合物制备了特异性的兔抗甲磷多抗血清。抗血清的ＥＬＩＳＡ效价为１：１２８００－１;５１２００,抗血清与乙酰甲胺磷的交叉反应率为１０．５％,与供试的其它９种有机磷农药的交叉反应率均小于２．８％,用同样的方法合成了甲胺磷－卵清蛋白复合物作为包被抗原,建立了适于环境中甲胺磷残留分析的间接竞争ＥＬＩＳＡ工程程序,在明确     

溴甲烷在土壤及作物（黄瓜）中的消解动态

用衍生气相色谱法测定了溴甲烷熏蒸大棚土壤后土壤及作物（黄瓜）中残留的Ｂｒ＾－，用顶空法测定土壤吸附的溴甲烷。在０．４－４０ｍｇ／ｋｇ的添加范围内，Ｂｒ＾－的平均回收率为８３．７％－９４．８％，相对标准偏差为±２．５％￣１４．７％。Ｂｒ＾－在土壤和黄瓜中的半衰期为１５．０和１４．２ｄ（北京）、２４．７和１０．３ｄ（沈阳）。土壤吸附的溴甲烷量较少，其半衰期和Ｂｒ＾－的相近。低于７５．０ｇ／ｍ＾２的施用     

柠檬桉组织培养的初步研究

本文初步研究了柠檬桉（ＥｕｃａｌｙｐｔｕｓｃｉｔｒｉｏｄｏｒａＨｏｏｋ．）茎段培养中腋芽伸长生长、继代增殖及试管苗生根的主要影响因素。在含有６－ＢＡ０．１～２．５ｍｇ／Ｌ、ＮＡＡ０．１～０．５ｍｇ／Ｌ的激素组合的改良ＭＳ培养基上都能诱导柠檬桉茎段腋芽伸长生长，最高诱导率可达７３％。继代增殖以６－ＢＡ０．５～１．０ｍｇ／Ｌ为宜，约２０～３０ｄ，丛生芽可增殖４左右，以每年７代计，年增殖率约１．６×１０４。继代培养基的６－ＢＡ浓度过高会导致丛生芽在形态上产生变异，并对生根产生不利的影响。继代增殖在７代内生活力未见有明显衰退。ＩＢＡ对柠檬桉试管苗生根有良好的促进作用，浓度以０．５～２．０ｍｇ／ｔ较为适宜。生根效果不仅与ＩＢＡ浓度有关，还与继代丛生芽的状态有关。生根的小苗已成功移栽到室内的盆中。本研究结果表明，可以用组织培养技术，快速繁殖经过选优的柠檬桉优良无性系。     

砂土供磷特性及磷肥效应研究

砂土供磷特性及磷肥效应的研究结果表明，砂土磷素形态以无机磷为主，有机磷甚少，仅占全磷含量的８．８４％。无机磷占９１．１５％，无机磷形态组成以磷酸钙为主，Ｃａ－Ｐ平均为无机磷总量的８６．１％，Ｏ－Ｐ占９．６％，Ａｌ－Ｐ占３％，Ｆｅ－Ｐ占１．３％。各种形态无机磷与速效磷的关系为：ｙ（速效磷）＝４９．３０ｘ１（ＡＬ－Ｐ）＋２１．８５４７ｘ４（Ｃａ－Ｐ）－３．４２０９，表明Ａｌ－Ｐ、Ｃａ－Ｐ对砂土速效磷的     

脑钠肽与谷胱甘肽巯基转移酶融合蛋白的表达与分离纯化

用ＰＣＲ技术扩增获得脑钠肽－９５ｃＤＮＡ,再由它获得脑钠肽－３８ｃＤＮＡ（ＢＮＰ－３８ｃＤＮＡ）选用融合蛋白表达质粒ｐＧＥＸ－３Ｘ作载体,构建成谷胱甘肽巯基转移酶（ＧＳＴ）和ＢＮＰ－３８的融合基因,该基因在大肠杆菌ＪＭ１０９中表达的最佳体系是：以１：１０扩大培养４ｈ加入终浓度为０．５ｍｍｏｌ／Ｌ的诱导剂ＩＰＴＧ诱导培养４ｈ,诱导表达的融合蛋白量可达５６μｇ／ｍＬ,培养液,是国外文献的３．７３倍,用     

晋中东山地区褐土土壤水分特征的测定与应用

利用压力膜方法测定晋中东山地区褐土的土壤水分，并分析了土壤水分及其能态特征。结果表明：本区土壤水吸力与土壤含水量之间呈明显的幂函数关系，可以用幂函数方程模拟；土壤的当量孔径分布大致随土壤水吸力的增加而减小；土壤的结构性能良好，通气孔隙、毛管孔隙、无效孔隙的当量孔径分布分别占土壤总孔隙度的１８．３％、５１．２％和３０．５％，尤其是毛管孔隙的分布占土壤总孔隙度的一半以上；土壤的比水容量随吸力的增大而迅速减小．当吸力为０．１×１０￣５～０．３×１０￣５ｐａ时，其比水容量的量级为１０￣（－１）［ｍ１／（１０￣５ｐａ·ｇ）］，而当吸力达３×１０￣５～１５×１０￣５ｐａ时，其比水容量的量级下降了１００倍，为１０￣（－３）［ｍｌ／（１０￣５Ｐａ·ｇ）］。表层土壤水分对于作物吸收利用比心土层和底土层更为有效，但深层土壤的持水性能比表层士壤要好。为此，必须提高作物对土壤深层储水的利用。     

高固氮基因工程大豆根瘤菌的研制及其增氮效应

通过构建快生型大豆根瘤菌Ｂ５２的基因文库和三亲本杂交，将增效因子ＤＮＡ片段导入优良的慢生型大豆根瘤菌２２－１０中，获得携带来自快生菌增效因子ＤＮＡ片段的工程菌株ＨＮ３２，经盆栽和小区试验，证明基因工程菌株ＨＮ３２比出发菌株２２－１０平均增产６％，比对照平均增产１３．２％～１６．９％，相当于每公顷施７５～１５０ｋｇ尿素．１９９２～１９９５年，在广西推广应用基因工程大豆根瘤菌ＨＮ３２２．１６万ｈｍ２，每公顷平均增产１９％，投入产出比１：３０。增加经济效益１４０９．８万元。     

菌根真菌对茶树吸收磷、钾素的影响

以辐照灭菌的酸性黄壤为供试土，种植有性繁殖的茶苗并接种ＶＡ菌根真菌，以不接种菌根真菌为对照，分别追施３２Ｐ－过磷酸钙和＾８６Ｒｂ－氯化铷，试验结果表明：菌根侵染率，＾３２Ｐ组合合为５２．６０％，＾８６Ｒｂ组合为５６．７０％，茶树株高分别对照的２．１０倍和２．５０倍，对磷、钾元素的吸收量分别比对照增加４．６０和３．１０倍，磷肥利用率提高１４．１０％，钾肥利用率提高１７．１３％。     

新疆南天山乌什谷地灌淤土及其在土壤系统分类中的地位

乌什谷地水旱轮作中的水耕作用是引起灌淤土成土过程和成土特征变异的主要动力，形成的人为土具有灌淤表层的全部诊断条件，其变异主要表现在：活性Ｆｅ２Ｏ３增多，耕层至心土层达４克／千克以上，占土壤全Ｆｅ２Ｏ３的１０％以上，心土层下呈减少趋势，铁的晶胶率耕层低，而心，底土中显著增高，Ｋｈ值（层段晶化度／表层晶化度）１．３－４．５之间，多数剖面有锈斑；耕下均有鳞片状结构，不具有双重熟化和双重淋淀层段，本文中将     

AFLP and RFLP linkage map in <Emphasis Type="Italic">Coix</Emphasis>

Coix is a genus in the grass family placed in the tribe Maydeae. It is closely related to maize and is also used as a crop plant. Since many valuable traits have been identified recently in Coix, it is considered to be a valuable genetic resource, particularly for maize improvement. In this study, a Coix genetic linkage map was constructed using an F2 population of 131 individuals. Eighty AFLP and 10 RFLP markers were mapped, covering a total length of 1339.5 cM with an average interval of 14.88 cM. The map consisted of 10 linkage groups, were consistent with the chromosome numbers observed cytogenetically. Both AFLP and RFLP markers were used first for genetic analysis in Coix. AFLP markers were generated by two restriction enzyme combinations, EcoRI/MseI and PstI/MseI. A total of 1349 bands were amplified, of which 140 were polymorphic. The polymorphism detection efficiency of the two enzyme combinations was compared, and utility of AFLP markers to construct the linkage map was discussed. Ten RFLP markers detected by three different probes were distributed on eight different linkage groups. The results provide a foundation to map and isolate important genes in Coix, and to investigate its genomic architecture, possible origins, and relationship with maize at the DNA level.     

Community structure of the microbiota associated with nodal roots of rice plants along with the growth stages: estimation by PCR-RFLP analysis

Community structure of the microbiota in rice roots that developed from different nodes and at different growth stages were compared by using PCR (polymerase chain reaction)-RFLP (restriction fragment length polymorphism) analysis of 16S rDNA. This was the first study to have applied a molecular microbial technique for elucidating the rhizospheric microbial succession of rice roots. Rice plants were grown in submerged soil pots, and nodal roots were collected 5 times during the growing period of rice plants. The RFLP fragments digested by four restriction enzymes (Hinf I, Hae III, Sau3A I, and EcoR I) tended to increase along with the growth stage. A marked increase in the RFLP fragments coincided with the development of reduction in the rhizosphere soil. RFLP fragments that were associated with every nodal root irrespective to the sampling date and those specific to the early and late growth stages were identified. Systematic changes in RFLP patterns from higher (younger) nodal roots to lower (older) nodal roots were also observed at each sampling date, which indicated the succession of the microbial community from higher to lower nodal roots. Cluster analysis divided the RFLP patterns of the microbial community associated with nodal roots into four clusters depending on the growth stages of rice plants. The cluster of the early growth stage was further divided into two subclusters of higher and lower nodal roots. The specific RFLP fragments that contributed to the seasonal variation of the microbial community associated with nodal roots as well as those that characterized the aging of nodal roots were clarified by principal component analysis.     

水稻光叶性状基因gl1的精细定位与候选基因分析

光叶水稻是一类在叶片和谷粒等表面都光滑无毛的水稻,是美国南部和非洲一些国家的主栽品种。已有研究表明水稻的光叶性状受一对位于第5染色体上的隐性基因gl1控制。本试验旨在对gl1进行精细定位,分析候选基因,为最终克隆gl1基因、研究毛状体形态建成的调控奠定基础。首先根据与gl1连锁的RFLP标记在第5染色体上的大致位置,有...     

Molecular phylogenetic relationships of puffer fish inferred from partial sequences of cytochrome b gene and restriction fragment length polymorphism analysis

Phylogenetic relationships among puffer fish were investigated by comparing cytochrome b gene sequences and restriction endonuclease assays of 16 species from Taiwan. DNA was prepared for sequencing by PCR. No variation in sequences was detected among individuals within each species. Direct estimates of mitochondrial cytochrome b gene sequence divergence among 16 puffer fish were from 3.41 to 31.78%. Different restriction patterns were found among 16 puffer fish with 10 restriction endonucleases, whereas no variation in patterns was detected among individuals within each species. The polymorphisms obtained by RFLP have provided a new set of genetic markers for the accurate identification of sibling puffer species. It is the first molecularly based study of puffer diversity and sheds light on the evolution and taxonomy of this major puffer fish family.     

Identification of the razor clam species Ensis arcuatus, E. siliqua, E. directus, E. macha, and Solen marginatus using PCR-RFLP analysis of the 5S rDNA region

Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis of the 5S ribosomal DNA region has been applied to the establishment of DNA-based molecular markers for the identification of five razor clam species: Ensis arcuatus, E. siliqua, E. directus, E. macha, and Solen marginatus. PCR amplifications were carried out using a pair of universal primers from the coding region of 5S rDNA. S. marginatus was simply distinguished by the different size of the amplicons obtained. Species-specific restriction endonuclease patterns were found with the enzymes Hae III for E. arcuatus, E. siliqua, and E. directus, and Acs I for E. macha, and when two enzymes were combined, the four species were also identified. Thus, this work provides a simple, reliable, and rapid protocol for the accurate identification of Ensis and Solen species in fresh and canned products, which is very useful for traceability and to enforce labeling regulations.     

Carotenoid biosynthetic pathway in the citrus genus: number of copies and phylogenetic diversity of seven genes

The first objective of this paper was to analyze the potential role of allelic variability of carotenoid biosynthetic genes in the interspecific diversity in carotenoid composition of Citrus juices. The second objective was to determine the number of copies for each of these genes. Seven carotenoid biosynthetic genes were analyzed using restriction fragment length polymorphism (RFLP) and simple sequence repeats (SSR) markers. RFLP analyses were performed with the genomic DNA obtained from 25 Citrus genotypes using several restriction enzymes. cDNA fragments of Psy, Pds, Zds, Lcy-b, Lcy-e, Hy-b, and Zep genes labeled with [alpha-(32)P]dCTP were used as probes. For SSR analyses, two primer pairs amplifying two SSR sequences identified from expressed sequence tags (ESTs) of Lcy-b and Hy-b genes were designed. The number of copies of the seven genes ranged from one for Lcy-b to three for Zds. The genetic diversity revealed by RFLP and SSR profiles was in agreement with the genetic diversity obtained from neutral molecular markers. Genetic interpretation of RFLP and SSR profiles of four genes (Psy1, Pds1, Lcy-b, and Lcy-e1) enabled us to make inferences on the phylogenetic origin of alleles for the major commercial citrus species. Moreover, the results of our analyses suggest that the allelic diversity observed at the locus of both of lycopene cyclase genes, Lcy-b and Lcy-e1, is associated with interspecific diversity in carotenoid accumulation in Citrus. The interspecific differences in carotenoid contents previously reported to be associated with other key steps catalyzed by PSY, HY-b, and ZEP were not linked to specific alleles at the corresponding loci.     

浙鲜9号的航天选育与特征特性分析

为探究航天诱变对大豆主要农艺性状的影响,本研究比较了航天诱变选育新品种浙鲜9号与其亲本台湾75在生育期、产量、品质、抗病性等方面的差异。结果表明,浙鲜9号播种至采收生育期比亲本短2 d,株高矮7 cm,鲜百荚重高6.1 g,鲜百粒重低2.2 g,青荚色比亲本淡,两年区域试验平均鲜荚产量较亲本显著增加9.4%。浙鲜9号不仅保留了亲本口感甜糯的优良品质,而且对大豆花叶病毒病的抗性有大幅度提高。利用60对核心SSR引物对二者进行分析,在Satt184、Satt197、Sat_-213等10个标记间发现多态性位点,引物多态性率为16.7%,Sat_-213为大豆花叶病毒病抗性基因Rsc15相关分子标记,这从分子水平证实了浙鲜9号抗性的改良。采用100个SNP标记对二者进行分析,有5个SNP标记在二者之间存在差异。浙鲜9号与亲本主要特征特性和DNA分子标记的对比研究均充分证明了航天诱变育种的有效性和可靠性。     

Development of Expressed Sequence Tag (EST)-based Cleaved Amplified Polymorphic Sequence (CAPS) markers of tea plant and their application to cultivar identification

To develop cleaved amplified polymorphic sequence (CAPS) markers for cultivar identification of the tea leaf, 5 primer pairs designed on the basis of genes that encode proteins related to nitrogen assimilation and 26 primer pairs based on expressed sequence tag (EST) sequences of the root of tea plant were screened. From combinations of primer pair and restriction enzyme that showed polymorphism among tea plants, 16 markers were selected and applied to DNA fingerprinting of Japanese tea cultivars. Sixty-three cultivars, except for a bud sport (Kiraka) and its original cultivar (Yabukita) and a pair that was the progeny of the same crossing parent (Harumoegi and Sakimidori), were distinguished from one another. By combining the 16 markers with previously developed CAPS markers and observing the physical appearance, 67 cultivars were distinguishable. The cultivars involve approximately 95% of total tea cultivating area in Japan; therefore, about 95% of tea leaves produced in Japan can be authenticated by labeling their cultivars.     

47份水稻品种资源的ISSR遗传多样性分析

为研究广东省惠州市种植的常规水稻品种的遗传多样性,本实验利用ISSR标记对47份水稻品种资源进行遗传多样性检测。从49条引物中筛选出5条重复性好,条带清晰的引物进行PCR扩增,共扩增出53条带,每个引物可以扩增出9～13条带,平均为10．6条,其中47条具有多态性,比率为88．7％。不同水稻品种间遗传相似系数变幅为0．319～0．936,平均达0．691,说明ISSR标记能够揭示材料间较高的遗传多样性。通过聚类,从分子水平对水稻品种资源的遗传关系进行分析,并对47份水稻品种资源进行分类,ISSR标记能将47份水稻品种完全区分开,为水稻品种资源的研究利用提供参考。     

Evidence of rapid evolution and incipient speciation in Vicia sativa species complex based on nuclear and organellar RFLPs and PCR analysis

Seventy three accessions of the sevenVicia species belonging to Sativa speciescomplex were screened for nuclear and organellar restriction fragmentlength polymorphic (RFLP), and random amplified polymorphicDNA (RAPD) markers. Total genomic DNAs of 73 accessions wasrestricted with three enzymes, and the restriction fragments werehybridized to the wheat rDNA probe pTa71 (containing 18S, 5.8Sand 25S rDNA genes, and spacers), and faba bean probes Ver6-5 (entire intergenic spacer flanked by small part of25S and 18S fragments) and Ver 18-6 (part of thecoding region of the gene and internal transcribed spacers). InXbaI digests, 16 repeat unit length classes in24 combinations were identified. Digestion withEcoRI and DraI gave2–4 and 1–3 fragments, respectively, with detectablehybridization to the probes, indicating the existence of internalXbaI sites. All the accessions produced 3.5 EcoRI fragment arising fromcoding region of the repeat unit. Four hundred and eighteen RAPDmarkers among 45 accessions were identified with 14 arbitrary10-base primers. The percentage of polymorphic bands withinspecies ranged from 20 in V.angustifolia to 98% inV. nigra. Both RFLP andRAPD markers were unable to assess the relationships betweenaccessions within species as there was often much closer resemblancesbetween certain accessions of different species rather than betweenaccessions within each taxon. This analysis supports the view basedon morphological, cytogenetical and crossability data that it is notpossible to classify Sativa species complex into a small finitenumber of taxa which are clearly circumscribed, and that the complexrepresents a unique case of rapid evolution and incipient speciation.A study of chloroplast and mitochondrial RFLPs was undertaken toanalyze phylogeny through maternal lineage. Chloroplast DNArestriction fragment patterns, using 13 restriction endonucleases,revealed 92.6 to 99% homology between the seven species.Twelve enzyme-probe combinations yielded identical fragmentpatterns for all the seven species. The molecular sizes of thechloroplast DNAs obtained were similar (121.5–123.5), indicating that they had all lost one of theinverted repeats. Total DNAs digested with three restriction enzymesand hybridized to six heterologous probes of mitochondrial originyielded monomorphic bands in five enzyme—probe combinationsacross all the 73 accessions. In other combinations as well,40–66 accessions yielded monomorphic profiles. The smallvariation in the remaining accessions was not species-specificsince the same profiles were present in more than one species. Theseresults i) strongly suggest that the seven species within thecomplex share a common ancestor, or direct lineage and, ii)indicate that these species should be relegated to a rank, perhaps ofsubspecies, within V.sativa species complex.