β—glucuronidase基因在水稻植株中的表达

采用花粉管通道法，将带β－ｇｌｕｃｕｒｏｎｉｄａｓｅ（ＧＵＳ）基因的ｐＢＩ１２１质粒ＤＮＡ导入栽培稻农垦５８。用荧光法检测ＧＵＳ活性，证明ＧＵＳ基因在水稻转基因植株中得到表达。Ｄｏｔ ｂｌｏｔ和Ｓｏｕｔｈｅｒｎ ｂｌｏｔ的结果表明经花粉管通道途径导入的外源ＤＮＡ整合到了转基因植株的基因组中，本文还对转基因植株后代的各种变异进行了讨论。     

水稻花发育相关MADS-box基因的克隆

以水稻幼穗总ＲＮＡ为模板,利用３′－ＲＡＣＥ克隆了８个接近于全长的水稻ＭＡＤＳ－ｂｏｘ 基因的ｃＤＮＡ．在ＧｅｎＢａｎｋ 的数据库中查询表明,其中２个ＦＤＲＭＡＤＳ１,ＦＤＲＭＡＤＳ２分别与已报道的水稻ＭＡＤＳ－ｂｏｘ 基因Ｏｓ－ＭＡＤＳ５和ＯｓＭＡＤＳ４５有极高的同源性,另外６个为新的尚未见报道的水稻ＭＡＤＳ－ｂｏｘ 基因．系统进化树分析表明ＦＤＲＭＡＤＳ１,ＦＤＲＭＡＤＳ２和ＦＤＲＭＡＤＳ４可能与Ｃ组基因的功能相关;ＦＤＲＭＡＤＳ３,ＦＤＲＭＡＤＳ６和ＦＤＲ－ＭＡＤＳ７可能与Ａ 组基因的功能相关     

水稻花发育相关MADS—box基因的克隆

以水稻幼穗总ＲＮＡ为模板,利用３’－ＲＡＣＥ克隆了８个接近于全长的水稻ＭＡＤＳ－ｂｏｘ基因ｃＤＮＡ。在ＧｅｎＢａｎｋ的数据库中查询表明,其中２个ＦＤＲＭＡＤＳ１,ＦＤＲＭＡＤ＃２分别与已报道的水稻ＭＡＤＳ－ｂｏｘ基因ＯＳＭＡＤＳ５和ＯｓＭＡＤ４５有极高的同源性。另外６个为新的未见报道的的水稻ＭＡＤＳ－ｂｏｘ基因。     

细茎大豆（G.gracilis）rbcS基因结构与分子进化分析

从细茎大豆的嫩叶中提取总ＤＮＡ，用ＰＣＲ方法扩增得到包含完整编码区的ｒｂｃＳ基因并将其克隆到ｐＢＬＵＥＳＣＲＩＰＴ载体中。完成全基因１０８９个核苷酸测序后，运用ＰＣＧＥＮＥ进行顺序编辑和同源比较，并应用ＭＥＧＡ１．０２１软件中的Ｎｅｉｇｈｂｏｒ－ｊｏｉｎｉｎｇ方面画出Ｒｕｂｉｓｃｏ小亚基仓的系统进化树。     

SURFACE PINNING AND ITS DETERMINATION BY MAGNETIC RELAXATION

ＳＵＲＦＡＣＥＰＩＮＮＩＮＧＡＮＤＩＴＳＤＥＴＥＲＭＩＮＡＴＩＯＮＢＹＭＡＧＮＥＴＩＣＲＥＬＡＸＡＴＩＯＮＺｅｎｇＺａｏｙａｎｇ１）ＤｉｎｇＳｈｉｙｉｎｇ１）ＹａｏＸｉｘｉａｎ１，２）（１）ＰｈｙｓｉｃｓＤｅｐａｒｔｍｅｎｔａｎｄＳｏｌｉｄＳｔａｔ...     

关于高校内部管理的四个标准

关于高校内部管理的四个标准ＯＮＦＯＵＲＣＲＩＴＥＲＩＡＯＦＴＨＥＩＮＴＥＲＩＯＲＭＡＮＡＧＥＭＥＮＴＯＦＴＨＥＵＮＩＶＥＲＳＩＴＩＥＳＡＮＤＣＯＬＬＥＧＥＳＣｈｅｎＺｈａｎｇｍｉｎｇ（ＺｈｅｊｉａｎｇＦｉｓｈｅｒｉｅｓＣｏｌｌｅｇｅ，Ｚｈｏｕｓｈａｎ...     

ESTABLISHING AN ORGANIC CHEMISTRY MICROSCALE LABORATORY PROGRAM

ＥＳＴＡＢＬＩＳＨＩＮＧＡＮＯＲＧＡＮＩＣＣＨＥＭＩＳＴＲＹＭＩＣＲＯＳＣＡＬＥＬＡＢＯＲＡＴＯＲＹＰＲＯＧＲＡＭ￥ＡｒｔｈｕｒＲ．Ｍｕｒｄｏｃｈ（ＭｏｕｎｔＵｎｉｏｎＣｏｌｌｅｇｅ，Ａｌｌｉａｎｃｅ，Ｏｈｉｏ，ＵＳＡ）（Ｖｉｓｉｔｉｎｇｐｒｏｆｅｓ...     

关于自然坐标系符号问题的讨论

关于自然坐标系符号问题的讨论龚礼宾（内江师专内江６４１００）ＳＯＭＥＣＯＮＳＩＤＥＲＡＴＩＯＮＳＡＢＯＵＴＴＨＥＳＩＧＮＳＩＮＴＨＥＮＡＴＵＲＡＬＣＯＯＲＤＩＮＡＴＥＧｏｎｇＬｉｂｉｎ（ＮｅｅｊｉａｎｇＴｅａｃｈｅｒｓＣｏｌｌｅｇｅ，Ｎｅｉｊｉａｎｇ...     

ACOUSTIC NONLINEARITY PARAMETER TOMOGRAPHY WITH FINITE AMPLITUDE SOUND WAVE

ＡＣＯＵＳＴＩＣＮＯＮＬＩＮＥＡＲＩＴＹＰＡＲＡＭＥＴＥＲＴＯＭＯＧＲＡＰＨＹＷＩＴＨＦＩＮＩＴＥＡＭＰＬＩＴＵＤＥＳＯＵＮＤＷＡＶＥＺｈａｎｇＤｏｎｇ；ＧｏｎｇＸｉｕｆｅｎ；ＹｅＳｈｉｇｏｎｇ（ＩｎｓｔｉｔｕｔｅｏｆＡｃｏｕｓｔｉｃｓ，ＫｅｙＬａｂ...     

SIMILARITY SOLUTION FOR A SNR EVOLVING IN AN INHOMOGENEOUS STELLAR WIND

ＳＩＭＩＬＡＲＩＴＹＳＯＬＵＴＩＯＮＦＯＲＡＳＮＲＥＶＯＬＶＩＮＧＩＮＡＮＩＮＨＯＭＯＧＥＮＥＯＵＳＳＴＥＬＬＡＲＷＩＮＤＣｈｅｎＹａｎｇ；ＬｉｕＮｉｎｇ；ＷａｎｇＺｈｅｎｒｕ（ＤｅｐａｒｔｍｅｎｔｏｆＡｓｔｒｏｎｏｍｙ，ＮａｎｊｉｎｇＵｎｉｖｅｒｓ...     

褐飞虱喂养试验显示表达GNA的转基因水稻纯系抗褐飞虱

利用基因枪法将含有3个不同基因（hpt,gus和gna）的质粒pWRG1515和pRSSGNA1共同转化粳稻品种鄂晚5号成熟胚诱导的愈伤组织。共再生出35株独立转基因植株。PCR／Southern印迹法分析发现,83％的转基因植株含有所有3个外源基因。Western印迹法分析发现79％的含gna基因的转基因植株以不同水平表达GNA。遗传分析证实外源基因在转基因植株后代中以孟德尔方式遗传。从其R1代亲本为孟德尔3：1方式遗传的R2代中,鉴定出2个含有所有3个外源基因的独立转基因植株纯系。这些纯系具有相似的外源基因表达量。褐飞虱喂养试验表明,这些纯系对褐飞虱具有显著的抑制作用。这些褐飞虱抗性提高的转基因纯系将应用于水稻抗虫育种中。实验证明,通过遗传转化和筛选可获得含在农业上有应用价值基因的转基因水稻纯系。     

Transgenic rice homozygous lines expressing GNA showed enhanced resistance to rice brown planthopper

Mature seed-derived calli from two elite Chinese japonica rice (Oryza sativa L.) cultivars Eyi 105 and Ewan 5 were co-transformed with two plasmids, pWRG1515 and pRSSGNA1, containing the selectable marker hygromycin phosphotransferase gene (hpt), the reporter β-glucuronidase gene (gusA) and the snowdrop (Galanthus nivalis) lectin gene (gna) via particle bombardment. 61 independent transgenic rice plants were regenerated from 329 bombarded calli. 79% transgenic plants contained all the three genes, revealed by PCR/Southern blot analysis. Western blot analysis revealed that 36 out of 48 gna-containing transgenic plants expressed GNA (75%) at various levels with the highest expression being approximately 0.5% of total soluble protein. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From the R2 generations whose R1 parent plants showing 3:1 Mendelian segregation patterns, we identified five independent homozygous lines containing and expressing all the three transgenes. Insect bioassay and feeding tests showed that these homozygous lines had significant inhibition to rice brown planthopper (Nilaparvata lugens, BPH) by decreasing BPH survival and overall fecundity, retarding BPH development and declining BPH feeding. These BPH-resistant lines have been incorporated into rice insect resistance breeding program. This is the first report that homozygous transgenic rice lines expressing GNA, developed by genetic transformation and through genetic analysis-based selection, conferred enhanced resistance to BPH, one of the most damaging insect pests in rice.     

Expression of two insect-resistant genes crylA ( b&c )/GNA in transgenic tobacco plants results in added protection against both cotton bollworm and aphids

The synthesized Bacillus thuringiensis insecticidal protein gene crylA(b&c) and the synthesized gene GNA, (the mannose specific lectin from snowdrop ( Galanthus nivalis)), tumefaciens have been inserted into plant expression vector pGW4BAI. Leave stripes of Nico-tiana tabacum var. K326 have been transformed with Agrobacterium tumefaciens strain LBA4404 harboring the plant expression vector. 28 kanamycin resistant tobacco plants have been obtained. PCR and Southern blot analyses show that the foreign crylA and GNA genes have been inserted into the genome of transformed tobacco plants. Haemagglutination assays show that GNA has a functional activity. Leaf disc bioassays against cotton bollworm ( H. armigera) show that the transgenic tobacco plants have a high insecticidal activity. The inhibition of aphid population in leaf disc bioassays against Myzus persicae shows that the fecundity of aphid on transgenic plants is lower than that on untransformed plants; the aphid population on the transgenic tobacco plants is 25%-70% that on untransformed tobacco plants. ELISA analysis of CrylA protein in tobcco leaves provides similar data to bioassay results. Through the two bioassays against H. armigera and M. persicae, several transgenic tobacco plants showing high insect-resistant activities to both pests have been obtained.     

将GNA基因导入莴苣（L.sativa）及其表达的研究

将含ＣａＭＶ３５Ｓ启动子的雪花莲凝集素基因置换Ｔｉ质粒载体ｐＢＩ１２１中的ＧＵＳ基因得到了ｐＢＩＧＮＡ质粒,将其导入农杆菌ＬＢＡ４４０４,得到ＬＢＡ４４０４菌株。     

抗除草剂旱稻转基因植株的获得

以旱稻丹粳旱５－５５、６－２４、６－３７的悬浮细胞及未成熟胚为受体材料，采用基因枪法将含ＢＡＲ基因的ｐＤＭ３０２质粒ＤＮＡ导入旱稻细胞中，经ＰＰＴ筛选获得抗性愈伤组织，筛选出的愈伤组织在分化培养基上再生出完整的旱稻转基因植株。Ｓｏｕｔｈｅｒｎ分子杂交分析表明，外源ＢＡＲ基因已整合到水稻基因组内，抗除草剂试验结果表明，转化植株对０．０１％Ｂａｓｔａ（有效成分为ＰＰＴ）有一定程度的抗性，说明外源ＢＡＲ     

Overexpression of glutamine synthetases confers transgenic rice herbicide resistance

Glutamine synthetase (GS, E.C.6.3.1.2) is a key enzyme involved in the assimilation of inorganic nitrogen in higher plants and gram-negative microorganisms. GS is the targeting enzyme of a herbicide phosphinothricin (PPT) or Basta. In order to generate PPT-resistant transgenic rice via overexpression of GS, we constructed a plant expression vector p2GS harboring two different isoenzymes GS1 and GS2 cDNAs under the control of constitutive promoters of rice Act1 and maize Ubiquitin(Ubi) genes. The p2GS was introduced into rice genome by Agrobacterium-mediated transformation and confirmed by PCR and Southern blot hybridization. GS-transgene expression was first detected by Northern blot analyses. Results from Basta test indicated that GS-transgenic plants can tolerate as high as 0.3% Basta solution. In addition, our results also demonstrated that GS overexpression conferred transformed rice calli PPT resistance. Thus, GS cassette can serve as a selective marker gene instead of bar cassette for selection of PPT transformants.     

表达雪花莲外源凝集素基因的油菜转基因植株的获得

利用农杆菌素LBA4404（pCAMBIA3300RG)转化优良甘蓝型油菜恢复系W723的下胚轴节段.pCAM-BIA3300RG含有Rssl启动子引导的雪花莲外源凝集素基因（gna）和CaMV-35S启动子引导的除草剂抗性基因（bar）。经过两轮除草剂（2.5mg/L bialaphos）筛选（两周/轮）,除草剂抗性再生芽被传入生根培养基中生根,对根系旺盛生长的植株中所含gna基因进行PCR分析,PCR分析证实了这些植株确为转基因植株,利用Western印迹法对随机选择的5株含gna基因的转基因植株的分析发现,其中4株表达了gna基因。目前正对这些表达gna基因的转基因植株进行后代遗传分离分析。     

Salt tolerance of transgenic rice (Oryza sativa L.) with mtlD gene and gutD gene

Southern blot analysis indicated that mtlD gene (encoding mannitol-1-phosphate dehydrogenase) and gutD gene (encoding glucitol-6-phosphate dehydrogenase) had been integrated into the rice genome mediated by Agrobacterium tumefaciens LBA4404(pBIGM). The expression of the above two genes in transgenic rice plants was demonstrated by Northern blot analysis and enzymatic activity assay. Analysis of sugar alcohol showed that transgenic rice plants could produce and accumulate mannitol and sorbitol. The salt tolerance of transgenic plants was much higher than that of their controls.     

Salt tolerance of transgenic rice (Oryza sativa L.) withmtlD gene andgutD gene

Southern blot analysis indicated thatmtlD gene (encoding mannitol-1-phosphate dehydrogenase) andgutD gene (encoding glucitol-6-phosphate dehydrogenase) had been integrated into the rice genome mediated byAgrobacterium tumefaciens LBA4404(pBIGM). The expression of the above two genes in transgenic rice plants was demonstrated by Northern blot analysis and enzymatic activity assay. Analysis of sugar alcohol showed that transgenic rice plants could produce and accumulate mannitol and sorbitol. The salt tolerance of transgenic plants was much higher than that of their controls.