共查询到19条相似文献，搜索用时 203 毫秒
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GNA基因表达载体的构建及遗传转化甘蔗 总被引：2，自引：0，他引：2
利用经人工改造的gna基因分别与Ubi、Rbcs启动子组合,构建抗虫植物表达载体pUNG和pRNG,通过农杆菌介导法导入甘蔗愈伤组织,经PPT筛选和PCR检测,获得Ubi-gna转基因植株124株,Rbcs-gna转基因植株35株。对部分转基因植株进行RT-PCR检测,初步证明外源基因已经整合到甘蔗基因组中,并得到有效表达;对RT-PCR阳性植株进行GNA(植物外源凝集素)活性测验,结果表明,甘蔗叶片表达的凝集素可以使健康公鸡的红细胞凝集,初步证明表达的GNA具有正常的生物学活性。 相似文献
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将包含2个抗虫基因sbk\[修饰后的Cry1A(c)\]和sck（修饰后的CpTI）的质粒载体pCDMARUBA Hyg转入农杆菌EHA105中, 感染南粳45的愈伤组织, 得到再生植株。用sbk和sck基因的引物进行PCR分析, 从97个再生植株中筛选出42个含有2个抗虫基因而没有潮霉素基因的阳性植株。通过Southern杂交发现, 42个阳性植株中具有1~5个外源基因拷贝的分别有23、 11、 5、2和1个单株。用RT PCR检测发现, 4个单拷贝株衍生的28个T3代植株外源基因能正常表达。人工饲喂二化螟幼虫试验也表明, 转基因植株的幼虫死亡率达94%~100%, 抗虫能力比对照显著提高。筛选出3个农艺性状优良的株系。 相似文献
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《中国油料作物学报》2017,(2)
利用无缝克隆技术把抗虫基因Cry8-like重组到pCAMBIA3301载体上,成功构建了含有抗草铵膦筛选标记的植物表达载体pCAMBIA3301-RSP-Cry8-like-nos。将该载体通过农杆菌介导法转入两个受体大豆品种吉农28和吉农42中,经PCR检测,吉农28和吉农42分别获得了17株和13株T_1阳性植株,36株和25株T_2阳性植株。T_2转基因植株Southern杂交结果显示,目的基因以单拷贝的形式整合到大豆基因组中。荧光定量PCR检测结果表明,Cry8-like在转基因植株幼根得到了表达,而非转化受体对照未检测到荧光信号。室内抗暗黑鳃金龟幼虫鉴定结果表明,转基因植株提高了抗暗黑鳃金龟幼虫的能力。 相似文献
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农杆菌介导的DnMADS2基因转化金钗石斛初步研究 总被引：1，自引：0，他引：1
为建立稳定有效的金钗石斛转化体系,进一步研究花发育相关基因的功能,为石斛兰分子育种提供基础,本研究以金钗石斛类原球茎（PLBs）为转化受体,通过农杆菌介导的方法将DnMADS2基因转入金钗石斛。结果表明：用20 mg/L潮霉素筛选4个月后获得抗性PLBs,对其进行HPT基因和DnMADS2基因的PCR检测表明,目的基因已成功整合到PLBs中。获得的转化PLBs在抗性培养基上分化为不定芽后,再经生根壮苗培养获得转化植株。对转化植株进行Southern检测,结果表明,DnMADS2基因已整合到5株金钗石斛转化植株基因组中。据此建立的稳定高效的金钗石斛转化体系和获得的转基因植株,将为DnMADS2基因功能的进一步研究和金钗石斛转基因育种奠定基础。 相似文献
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通过基因枪(PDS-1000He)轰击法将构建好的多功能蛋白HC-Pro的植物表达载体pHCFL与带PMI筛选标记基因的pZMLR14植物表达载体共转化导入易感花叶病甘蔗品种福农95-1702的胚性愈伤组织中,转化的愈伤经甘露糖筛选后获得抗性转化甘蔗幼苗。经PCR检测、氯酚红显色、RT-PCR检测和DNA点杂交分析获得转HC-Pro的转基因植株,转化率为0.8%(HC-Pro基因)、1.7%(PMI基因)和0.4%(HC-Pro基因+PMI基因)。花叶病毒接种实验结果显示,未转基因植株的发病率为75%,而转基因植株均系统性发病,推测由于HC-Pro的大量表达抑制了甘蔗体内自身的RNA沉默机制而使病情加重。其它农艺性状调查结果显示,转基因与未转基因植株之间不存在显著差异,表明转HC-Pro和PMI基因对甘蔗的生长或其他农艺性状没有负面影响。 相似文献
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通过根癌农杆菌介导法,以子叶节为外植体将抗虫基因cryIA转入东农50大豆品种中,对获得的T0、T1和T2代植株采用PCR、RT-PCR及Southern杂交法进行分子检测,并对转基因大豆进行食心虫室内抗性鉴定。经PCR鉴定,T0植株有4株为阳性植株,T1有3株为阳性植株,T2有9株为阳性植株。对3株T1 阳性植株进行RT-PCR检测,均扩增得到1 800bp目标片段, Southern杂交分析显示,有2株出现杂交信号,分别为单拷贝和双拷贝。室内抗虫鉴定表明转基因植株食心虫抗性明显优于对照品种,在蛋白质及脂肪酸含量等品质性状上,与对照没有显著差异。证明cryIA基因已整合到受体大豆基因组中,该基因在大豆T1转基因植株中能够正常转录,抗虫基因cryIA的导入可以提高东农50对大豆食心虫的抗性,其后代的遗传稳定性还有待于进一步的研究。
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甘蔗抗旱生理测验及BADH基因PCR扩增的研究 总被引：2，自引：0，他引：2
对41份甘蔗种质材料进行了抗旱相关基因BADH(甜菜碱醛脱氢酶)基因PCR检测。结果显示，仅7份野生祖亲种种质材料及4份栽培品种材料有扩增带，且不同种质之间扩增带序列长度有差异。同时对30份甘蔗种质材料的离体叶片保水率、叶片相对含水量、气孔开度等水分生理指标进行测验，其结果与生产实际抗旱性表现基本一致。初步筛选出具有较强抗旱性生理和遗传基础的甘蔗祖亲种质和栽培品种种质材料。 相似文献
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Takeo Sakaigaichi Yoshifumi Terajima Makoto Matsuoka Shin Irei Seiji Fukuhara Takayuki Mitsunaga 《Plant Production Science》2013,16(2):327-332
ABSTRACT Sugarcane smut, caused by Sporisorium scitamineum, is one of the most important sugarcane diseases in Japan. Wild sugarcane, Saccharum spontaneum, is known to be a key breeding material to obtain high-yielding clones. In this study, we sought to identify Japanese wild sugarcane accessions with high resistance to smut. Thirty wild sugarcanes and three sugarcane cultivars were tested by the pinprick method. The results of the inoculation tests aided in identifying wild sugarcanes with high resistance to smut disease, namely JW90, Iriomote8, and Iriomote15. After screening the germplasm, progeny distribution of smut resistance from the inoculation test and dry matter productivity in the smut disease-free field were compared. The highly resistant wild sugarcane accession had a much better impact on progeny distribution of smut resistance compared with the susceptible accession. No relationship was found between smut resistance and dry matter productivity in both populations. 相似文献
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向大豆导入几丁质酶基因的初步研究 总被引：32，自引：8，他引：24
以根癌农杆菌的Ｔｉ质粒为介导，将抗真菌病的几丁质酶基因，导入默经江省栽培大豆一东农３７号，吉林２８号等１４个品种。从子叶节和下胚轴诱导出丛生芽，并再生植株，经ＰＣＲ和ＤＮＡ分子杂交检测，经ＰＣＲ和ＤＮＡ分子杂交检测的１１３株大豆幼苗中，有７株呈阳性反应，证明几丁质酶基因已导入并整合到大豆的基因组中。 相似文献
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探讨切段式联合收割机收获的甘蔗茎段糖分变化情况,为今后糖厂合理安排机械砍收顺序和入榨时间提供参考。分别对两个甘蔗品种粤糖94-128和粤糖00-236采取人工收获和机械化收获,分析其放置过程中（5 d内,自然条件下）蔗糖分、还原糖、重力纯度变化情况。结果表明：与人工收获方式相对比,两个品种在机械化切段式收获方式下的蔗糖分、还原糖、重力纯度在收获当天转化不显著,而从收获后第二天开始,糖分开始变化,具体为蔗糖分、重力纯度下降,还原糖分升高,粤糖00-236的糖分转化损失速度大于粤糖94-128。因此建议切段式收获的甘蔗茎段应该安排当天入榨,而在同一天砍收这两个品种的情况下应该优先安排粤糖00-236入榨。 相似文献
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基因枪介导甘蔗遗传转化研究初报 总被引：4，自引：0，他引：4
将从灰树花（Grifola frondosa）中分离克隆到的海藻糖合酶cDNA以正向插入植物表达载体pBI121/BamHI Sst1位点，获得一植物表达载体pBT；又用Ubi-L启动子插入植物表达载体pBI121/HindⅢ＋Xbal位点，获得重组质粒pUB,再把海藻糖合酶cDNA正向插入pUB/BanHI SstI位点，获得另一植物表达载体pUBT。然后通过基因枪法分别用植物表达载体pBT和pUBT转化甘蔗。再生植株的点杂交和PCR－Southern杂交表明海藻糖合酶基因已整合进甘蔗的基因组中。 相似文献
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通过农杆菌介导将菜豆几丁质酶基因导入马铃薯 总被引：10，自引：2，他引：8
选用两个生产上主栽马铃薯品种“鲁引 1号”和“鲁引 4号”的试管微型薯为材料 ,通过农杆菌介导成功地将菜豆几丁质酶基因导入到马铃薯中。试管微型薯薯片与农杆菌共培养 3d后 ,转到含卡那霉素 10 0mg/L的分化培养基上诱导不定芽分化。待抗性芽长到 1 0～ 1 5cm高时 ,转入含卡那霉素 10 0mg/L的液体培养基中进行生根筛选。经过分化和生根两轮筛选的转化植株的PCR检测结果均为阳性 ,而未经转化的对照植株为阴性 ,证明该筛选系统是可靠的。早熟品种鲁引 1号的转化频率明显高于晚熟品种克新 4号 ,其最高转化频率为 4 1 0 % ,平均每个外植体可得到 3 85株转化苗。转化植株的抗病性鉴定正在进行中 相似文献
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Takeo Sakaigaichi Yoshifumi Terajima Makoto Matsuoka Takayoshi Terauchi Taiichiro Hattori Shoko Ishikawa 《Plant Production Science》2016,19(2):323-329
Modern sugarcane cultivars are derived from the interspecific crossing between Saccharum officinarum and wild sugarcane, Saccharum spontaneum. The introgression of valuable characteristics from wild sugarcane is recognized as extremely important, but this process typically requires long-term effort over multiple generations of backcrosses owing to the low sugar content of the initial interspecific hybrids. In this study, we aimed to identify Japanese wild sugarcanes with high juice brix in order to promote effective interspecific crossing of sugarcane. Sixty-four accessions from the Nansei Islands and 70 accessions from the Honshu were evaluated for juice brix. Wild sugarcanes with high juice brix were demonstrated to exist among wild sugarcanes indigenous to the Honshu. A significant difference was observed between the median juice brix values of wild sugarcanes of the Nansei Islands and those of the Honshu. The relationship between juice brix and stem traits was then examined in 20 wild sugarcanes, 10 each from the Nansei Islands and the Honshu. The reproducibility of juice brix value in both experiments was confirmed. In contrast to juice brix, stem traits, such as length, diameter, and volume, were typically smaller in wild sugarcanes from the Honshu. Moreover, a negative correlation was observed between the index of stem volume and juice brix. In this study, we identified outstanding wild sugarcanes with high juice brix. Using germplasms from the identified wild sugarcanes in interspecific crossing could contribute to the increases in both yield and sugar content. 相似文献
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Utilization of exotic and diverse germplasm is needed to enhance the genetic diversity of cultivars. Genetically diverse lines provide ample opportunity to create favorable gene combinations, and the probability of producing a unique genotype increases in proportion to the number of genes by which the parents differ. Representative core collections (10% of the entire collection) have been suggested as a means to identify useful parents for crop improvement programs. The chickpea core collection (1956 accessions) was evaluated for 14 agronomic traits in two seasons to identify diverse agronomically superior chickpea germplasm. Season (year) and genotypic effects were significant for 13 of the 14 traits, while genotype × season effect was significant for 8 traits. The desi, kabuli, and intermediate type chickpeas differed significantly for days to maturity, basal secondary branches, pods per plant, seed yield, and 100-seed weight. In comparison to controls, 12 accessions flowered early, 15 produced greater seed yield, and 29 had greater 100-seed weight. Based on days to 50% flowering, pods per plant, seed yield, and 100-seed weight, 19 desi, 15 kabuli and 5 intermediate type chickpea germplasm lines originating from 10 countries were selected. The selected desi accessions produced 8.5% more seed yield and had 32% larger seeds than the control cultivar Annigeri while the selected kabuli accessions yielded at par with control L 550 but had 84% larger seeds. The 39 selected accessions and two control cultivars (Annigeri and L 550) were grouped by their first five principal components (PCs) into three clusters. Cluster 1 consisted of early maturing large-seeded kabuli types, cluster 2 early and late maturing desi types, and cluster 3 late maturing intermediate and kabuli types. Clusters 2 and 3 accessions had small to medium sized seeds. These accessions can be used in chickpea breeding programs to develop high yielding desi and kabuli cultivars with a broad genetic base. 相似文献