抗除草剂基因导入早稻（Oryza sativa）栽培品种

以生产上优良旱稻(Oryza sativa L．)新品种旱稻297、旱稻10号等的幼胚愈伤组织为转化受体材料,用基因枪法把抗Basta除草剂的bar基因导入了这些品种的愈伤组织,经两轮Basta抗性筛选和分化获得了再生植株,对再生植株进行PCR扩增和Southern杂交检测,T0和T1代Basta抗性实验表明,bar基因已整合到旱稻的基因组DNA中,并在T1代继续表达。对各品种幼胚培养的诱导、分化培养基实验表明,MB和MS培养基可作为这5个品种的愈伤组织诱导培养基,改良的RMB2、RMS2培养基可显著地提高愈伤组织的分化频率。实验所获得的转基因植株和建立的遗传转化系统。为早稻的抗除草剂分子育种和其它基因转化奠定了初步的基础。     

粳稻云引抗稻瘟病基因的遗传分析及其定位

摘要：用8个稻瘟病菌[Pyricularia grisea (Cooke) Sacc.]系对稻瘟病抗性材料云引进行田间注射接种鉴定。结果表明，云引对接种的8个菌系均表现为抗病反应。用抗病材料云引和感病材料丽江新团黑谷作亲本杂交获得F1，F1自交获得F2群体，用上述8个菌系分别对亲本、F1和F2群体进行田间注射接种，鉴定结果表明云引对所有8个菌系的抗性均表现为单基因控制的分离特性（抗感分离比为3∶1）。利用微卫星标记将云引对四川-43菌系的抗性基因初步定位于水稻的第11号染色体上，与标记RM202的遗传距离为3.8 cM。     

中外不同猪品种生长激素基因遗传多态性检测

成熟的猪生长激素(pGH)分子由190个氨基酸组成,分子量为22kD。1987年Vize等^[1]成功地获得了猪的GH基因,该基因全长为2231bp,由5个外显子和4个内含子构成。Thomoson等^[2]将pGH基因定位于12号染色体短臂1区1带。Yerle等^[3]用高分辨G带染色体原位杂交将pGH基因定位于     

旱稻(Oryza sativa)/稗草(Echinochloa caudata)//高梁(Sorghum bicolor)三属杂种表现及分析

以旱稻基因型远F2 1 [旱 65 (Oryzasativa) 长芒稗 (Echinochloacaudata) ]为母本 ,高粱(Sorghumbicolor)基因型沈农 1 33为父本 ,进行属间远缘杂交 ,获得三属间杂交实粒种子。对杂交种F0 、F1、F2 代连续观察结果表明 :( 1 )杂交F0 代高度不孕 ,并伴有杂交种发育夭折现象发生 ,结实率仅为 2 63‰ ;( 2 )F1杂种优势明显 ,株高、穗长、1级枝梗数、小花数、结实粒数、结实率、千粒重诸性状超母本优势为 0 61 %～ 2 0 5 6% ;主茎叶片数多于母本 1片 ,总叶面积超亲优势为 1 1 95 % ;F1出现母本没有的红芒、红护颖和紫柱头 3个可遗传性状 ;( 3)F2 除在生育期、株高及穗长等性状发生分离外 ,其典型特征是穗型、芒性、芒色、柱头颜色、护颖颜色等性状发生分离     

水稻（Oryza sativa L．）苹果酸酶（OsNADP-ME3）基因在逆境下的表达特性研究

水稻苹果酸酶（NADP-ME）是多基因家族,由3个胞质型NADP-ME和1个质体型NADP—ME构成。本研究针对水稻胞质型成员（命名为NADP-ME3）（NM001061367）进行初步的功能解析。克隆获得的NADP-ME3基因的cDNA序列全长为2240bp,其中5’非翻译区为151bp,3’非翻译区为376bp,开放读码框（ORF）长1713bp,编码570个氨基酸。为研究NADP-ME3在逆境胁迫下的表达量变化,No．hemblot检测结果显示,在NaCl、NaHCO,和PEG胁迫条件下,NADP-ME3随胁迫处理时间的不同表达量呈现不同程度的变化,推断NADP-ME3可能与非生物胁迫有应答关系,将NADP-ME3转入拟南芥中并通过观察转基因拟南芥在非生物胁迫下表型变化,发现NADP-ME3能够在一定程度上提高植物对非生物胁迫的耐受性。     

将新的人工雄性不育基因导入小麦栽培品种的研究初报

利用ＰＤＳ－１０００型氢气基因枪将我们构建的雄性不育基因导入小麦栽培品种的幼胚愈伤组织,经含除草剂Ｂａｓｔａ的抗性培养基分化和生根筛选后得到２７株绿苗,ＰＣＲ和Ｓｏｕｒｈｔｅｍ检测均发现有３种株呈阳性,初步表明基因已整合到小麦的基因组中,转化频率达０．２％左右。     

反义乙烯受体LeETR2基因对番茄的转化和功能分析

摘要：采用根癌农杆菌（Agrobacterium tumefaciens）介导法将LeETR2的部分反义序列导入番茄(Lycopersicon esculentum)子叶外植体，经抗性筛选和组织培养获得再生植株。通过对转基因植株中插入序列的PCR检测和Southern杂交，证明转化番茄成功。通过对LeETR2的功能的初步研究结果表明, 转基因植株顶端优势丧失、极端矮化和番茄果实的内源乙烯合成量增加。这提示此基因在植物乙烯信号感受与转导系统中起着负调控的作用。     

转抗虫基因彩色棉的获得

彩色棉以其绿色环保特性，深受人们的喜爱，但彩色棉的抗虫性差将影响到彩色棉品种的大面积种植推广，因此选育抗虫新品种，具有重要的意义。本研究采用改造的全长雪花莲凝集素基因GNAmm与合成的带有分泌信号肽编码序列的Bt crylAc基因，分别与韧皮部特异表达启动子SPP2P和组成型     

牛GlyCAM1基因遗传多态性的研究

摘要：以中国荷斯坦牛、鲁西黄牛、渤海黑牛为研究对象,利用CRS-PCR、PCR-SSCP及DNA测序技术检测了GlyCAM1基因的遗传多态性。结果表明：本研究首次发现在牛GlyCAM1基因外显子3的第2081（A/C）和内含子3的2417（C/T）位点存在突变,两位点在3个牛群中的等位基因频率A/B分别为0.7525/0.2475、0.6112/0.3888、0.3375/0.6625; 0.9046/0.0954、0.8383/0.1617、0.7875/0.2125;经过χ2适合性检验, 3种牛群在内含子3的突变达到Hardy-Weinberg平衡状态（P>0.05）,在外显子3鲁西黄牛和渤海黑牛达到Hardy-Weinberg平衡状态（P>0.05）,但中国荷斯坦牛的突变在此位点未达到Hardy-Weinberg平衡状态（P<0.05）。     

含粘质沙雷氏菌几丁质酶SchiA基因的植物转化质粒pBG1112构建和水稻遗传转化

摘要:将一个2.8 kb的CaMV35S启动子/SchiA编码区/Nos终止区融合基因插入到植物转化载体pCAMBIA1301的多克隆酶切位点，得到1个14.6 kb的新植物转化质粒pBG1112,用花器介导法转化水稻（Oryza sativa ），经PCR检测，初步证实已将目的基因整合到受体植物的基因组中。一部分转基因T3代潮霉素抗性阳性植株对水稻纹枯病(Rhizoctonia solani )和稻瘟病（Pyricularia oryzae）的抗性较非转化对照增强。RT-PCR表明抗病性植株为阳性，而不抗病的植株为阴性。将RT-PCR产物测序后，用BLAST软件分析序列可知，该序列为细菌几丁质酶基因核苷酸序列而不是植物几丁质酶基因的核苷酸序列。T4代转基因水稻的几丁质酶活力高于对照未转基因植株，表明转入的外源几丁质酶基因能正常表达。     

Genetic Erosion from Modern Varieties into Traditional Upland Rice Cultivars (Oryza sativa L.) in Northern Thailand

The purpose of this study was to assess the extent of genetic erosion of traditional upland germplasm in northern Thailand as a result of gene-flow from distinct strains carrying different genotypes. Even modern variety specific markers have not been developed, there is a comparative population in Laos. Thus, both populations were compared with various characters to evaluate gene-flow from modern variety to landraces. Glutinous and glabrous strains are predominated in Laos. However, such strains were drastically decreased in north–east Thailand. Gene diversity is higher in Thailand, compared to Laos at seven isozyme loci. This was a result of the higher frequencies of Indica strains and heterozygotes in Thailand. Plastid type was also determined by using an INDEL marker. Nearly half of Indica strains carried the Japonica plastid. Heterozygotes also tended to carry Japonica cytoplasm. Such nuclear–cytoplasm substituted strains and heterozygotes were probably generated by natural hybridization. Japonica strains tended to be a maternal donor rather than Indica ones. Or Indica strains would easily release pollens, which grow outside of upland fields.     

Integrated Nutrient-Management Technology for Direct-Seeded Upland Rice (Oryza sativa) in Northwestern Himalayas

Alarming climate change, rainfed upland farming, and low resource-use efficiency of conventional fertilizer management practices are major production constraints detrimental to rice productivity in the northwestern (NW) Himalayas. Recent agronomic intervention of direct-seeded rice (DSR) coupled with suitable rice germplasm well suited to rainfed upland ecosystems in combination with appropriate integrated nutrient-management (INM) technology can enhance the rice productivity in the region. Thus, a field experiment with seven treatments replicated three times in a randomized block design was conducted on INM technology in rainfed upland rice cv. HPR-1156 (Sukaradhan-1) to harness the potential of DSR technology in order to boost rice productivity in the NW Himalayas. Results on INM in direct-seeded upland rice revealed that nitrogen, phosphorus, and potassium (NPK) at 90:45:45 kg ha^?1 + farm yard manure (FYM) at 5 t ha^?1 (oven dry-weight basis) significantly resulted in the greatest magnitude of growth and development (plant height, tillers m^?2) and yield-contributing characters (panicles m^?2, panicle length, grains panicle^?1 and 1000-grain weight), resulting in significantly greatest grain, straw, and biological yield followed by sole use of NPK at 90:45:45 kg ha^?1 and NPK at 60:30:30 kg ha^?1 + FYM at 5 t ha^?1, respectively, in rainfed upland rice. Application of NPK at 90:45:45 kg ha^?1 + FYM at 5 t ha^?1 again resulted in significant improvement in soil organic carbon and available NPK status over other treatments and initial soil fertility status in an acidic Alfisol. Overall, it is inferred that INM technology with judicious use of NPK at 90:45:45 kg ha^?1 + FYM at 5 t ha^?1 in rainfed upland rice under DSR technology can enhance the rice productivity and resource-use efficiency in NW Himalayas.     

Policosanol Content and Composition of Korean Rice (Oryza sativa L.) Cultivars

Policosanols (PCs) are a group of long‐chain alcohols that have been reported to have many beneficial physiological activities. In this study, the total content and composition of PCs in 15 rice (Oryza sativa L.) cultivars from Korea were characterized by gas chromatography–time‐of‐flight mass spectrometry. This method proved to be sufficiently precise and accurate with respect to the degree of endogenous biological variability found in the rice samples. Octacosanol (C28) and triacontanol (C30) were the major components of PCs in all cultivars. In addition, there were positive correlations among the determined PC contents. Given its high PC content, the Heughyangbyeo cultivar may appear to be a good candidate for future breeding programs.     

水稻密码子优化基因Mat#增强草铵膦抗性

来自细菌Nocardia sp.AB2253的蛋氨酸砜N-乙酰基转移酶基因(methionine sulfone Nacetyltransferase gene,Mat)编码产物具有N-乙酰基转移酶活性,能解除灭生性除草剂草铵膦的毒性,但在植物中表达效率较低.本研究用水稻(Oryza sativa)偏爱密码子优化的Mat#基因转化籼稻品系9K(Oryza sativa ssp.indica),经过PCR和Southern杂交验证,证明该基因已经整合至水稻基因组中.除草剂抗性检测结果显示,该转化体的芽期草铵膦耐受浓度至少为600 mg/L、秧苗期草铵膦耐受浓度至少为1 000 mg/L,对草铵膦的抗性水平不低于转双丙氨酰膦抗性基因(bialaphos resistance gene,Bar)水稻9KA2.酶活性测定结果显示,该转化体叶片中的N-乙酰基转移酶活性约为非转基因对照中的6.6倍.说明优化后的Mat#基因增强了草铵膦抗性,可作为转化筛选标记和抗除草剂目的基因应用于转基因作物育种.     

水稻簇生穗控制基因ts的定位及育种利用研究

从水稻(Oryza sativa L.)育种材料广恢128、粳稻Kitaake、蜀恢955和蜀恢881的复合杂交F2群体中发现了一个簇生穗突变体,该突变体命名为TS(top-spikelet-two-grain mutant)。TS最显著的变异为穗部一、二次枝梗顶端表现为2~3个小穗簇生在一起。为了明确TS中所含簇生基因ts的遗传机理及其在水稻育种中的利用价值,本研究利用TS与G2480B(indica)杂交的F1、F2群体进行簇生性状的形态观察和遗传分析;利用3个携带有ts基因的转育纯系分析ts基因的应用前景。结果表明,F1群体表现为全部显性,F2群体出现3∶1显隐性状分离,突变性状受1对隐性单基因控制,能够稳定遗传。采用简单序列重复(simple sequence repeat,SSR)标记技术,将突变基因ts定位于第6染色体上的长臂端,位于RM20323和RM6298之间,遗传距离分别为5.1和5.6 cM。3个转育纯系中,L332在保持簇生性状的同时,与亲本G2480B相比,其产量性状无显著差异,蒸煮食味品质较优。ts基因的表达与水稻的单株穗数、每穗粒数、结实率、千粒重和单株产量无显著相关性。以上结果为ts基因在水稻育种上的利用提供了一定的理论基础。     

OsEXPA8基因对水稻悬浮细胞的周期和大小的影响

膨胀素是植物细胞特有的一种细胞壁松弛蛋白,在很大程度上决定着植物细胞的生长速率.α-膨胀素(α-expansin,EXPA)和β-膨胀素(β-expansin,EXPB)是植物膨胀素家族中两个较大的亚家族.水稻(Oryza sativa)中至少有34个α-膨胀素基因.本研究通过转基因技术,研究了一个水稻根特异性表达的α-膨胀素基因OsEXPA8对水稻悬浮细胞的细胞周期和细胞大小的影响.利用流式细胞仪,分析不同培养时期野生型和转基因型细胞周期的差异,结果发现,35S∷OsEXPA8(过表达)转基因悬浮细胞大部分处于G2/M期,而OsEXPA8-RNAi (knock-down)转基因悬浮细胞多处在G1期;利用血球计数板测定细胞数量,结果表明,35S∷OsEXPA8转基因细胞数量＞野生型细胞数量＞OsEXPA8-RNAi转基因细胞数量;利用显微镜测定不同培养时期的细胞大小,发现35S∷OsEXPA8转基因细胞大小＞野生型细胞大小＞OsEXPA8-RNAi转基因细胞大小;测定愈伤组织块大小,发现35S∷ OsEXPA8转基因愈伤组织块大小＞野生型愈伤组织块大小＞ OsEXPA8-RNAi转基因愈伤组织块大小.本研究结果表明,OsEXPA8能够促进水稻悬浮细胞的分裂和生长,从而增加细胞数量,增大细胞体积,对水稻悬浮细胞的生长具有调控作用.该研究将有助于更好地了解植物细胞壁的松弛过程,从而深入地认识植物的生长发育进程.同时,膨胀素作为一种细胞壁松弛因子在农业上具有潜在的生物工程应用价值.     

我国转基因抗除草剂水稻的生态风险与控制

转基因抗除草剂水稻(Oryza sativa)的生态风险备受业界和公众关注。探讨其生态风险大小和收益多少,对于防范风险、明辨是非、权衡利弊、科学决策具有重要参考价值。以往研究注重单项风险分析,主要采用模拟条件分析,风险和效益的对比少、全局和集成分析不足。本文较全面地分析了转基因抗除草剂水稻的生态风险和收益,提出了控制生态风险的技术措施,对于促进我国转基因抗除草剂水稻的健康稳步发展具有现实意义。     

DDT and HCH Residues in Basmati Rice (Oryza sativa) Cultivated in Dehradun (India)

Organochlorine pesticides were used earlier for agricultureproduction. Their residues may still be present in soil and mayaccumulate in food crops, posing potential health problems to consumers. DDT, HCH, their isomers and metabolites were analyzedin samples of soil and rice plants collected from ten differentvillages of a well-known Basmati rice growing area in Dehradun.Residues of both pesticides were found in all samples ofsoil and different parts of rice plants except for a few grainsamples. Maximum residue was observed in husk and minimum ingrains. The average concentration of DDT in soil ranged from0.013 to 0.238 ppm. p,p′-DDE was the major metabolite (>63%). Theaverage concentration of DDT in rice grain varied from 0.002 to 0.040 ppm. o,p′-DDT was the main isomer (>93%). Theaverage concentration of HCH in soil ranged from 0.122 to 0.638 ppm. β-HCH was the predominant (43%) isomerfollowed by α-HCH (21%). The average HCH concentrationin rice grain ranged between 0.013 and 0.113 ppm. All four isomers were present in grains. The levels of DDT and CHCin grains were similar in magnitude as those from differentIndian states, but well below the maximum residue limit of 0.1 ppm for DDT and 0.05 ppm for HCH prescribed by the Government ofIndia and WHO/FAO. As such, the pesticide residue levels in thisexport commodity are not of hazardous nature.