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1.
《分子植物育种》2020,(14)
本研究以转Bt基因‘南林895’杨12个株系扦插苗为试验材料,分析其对美国白蛾和杨小舟蛾的虫害抗性。利用PCR、ELISA等技术对转抗虫基因杨树进行分子检测,同时在室内控制条件下对美国白蛾和杨小舟蛾进行抗虫性分析。PCR分析结果显示在12个转基因株系中均检测到了Bt目的基因片段。ELISA分析结果显示Bt毒蛋白浓度范围为371.9～10 698.2 ng/g,且不同株系间存在一定差异。转Bt基因杨树饲虫分析结果表明,12个转基因株系对美国白蛾和杨小舟蛾均有一定的杀虫活性,其中转基因株系A-4-6、A-5-0、A-3-4、A-5-23、Z-1-3的抗虫性较显著。株系A-5-0对杨小舟蛾1龄幼虫12 d校正死亡率达87.2%,对美国白蛾的18 d校正死亡率为65.6%。且转Bt基因‘南林895’杨对美国白蛾和杨小舟蛾幼虫的取食和生长有明显的抑制作用。在对12个转Bt基因株系的分子检测和饲虫试验中,Bt毒蛋白的表达水平存在差异且抗虫效果与Bt毒蛋白浓度的高低有关。 相似文献
2.
转双价抗虫基因BmkIT-Chitinase玉米株系的获得 总被引：2，自引：0，他引：2
通过超声波辅助花粉介导法,将双价抗虫基因BmkIT-Chitinase分别导入以玉米自交系昌7-2及郑58的花粉为受体的不同基因型的自交系中。本研究共处理玉米雌穗1072穗,获得T0代种子1563粒,经卡那霉素初筛,T1代～T4代PCR及SouthernBlot杂交分子跟踪检测共获得20个转化株系,田间抗虫性鉴定表明共有16个转化株系与对照在抗虫性方面有显著差异,且此抗性随着各代稳定遗传。农艺性状调查结果表明,所获得的转基因玉米株系中大部分材料的农艺性状与对照无显著差异,除了N55材料及N20-1材料。N55材料的穗位高度与对照相比略低6±0.5cm,而穗粒数增加75±5粒。而N20-1材料百粒重增加5±0.5g。因此,转入此双价抗虫基因对玉米农艺性状影响不是很大。经过分子检测、田间抗虫性鉴定及农艺性状调查我们最终选育了9个转双价抗虫基因昌7-2自交系优良株系,6个郑58转双价抗虫基因自交系优良株系。 相似文献
3.
转Bt+Sck基因双价抗虫棉的抗虫性及遗传分析 总被引：4，自引：1，他引：3
对通过花粉管通道注射获得的转Bt Sck双价基因抗虫棉纯系312-5T2和332-2T2进行了生物抗虫性测定,转基因纯系对棉铃虫的抗性显著高于非抗虫对照苏棉16和抗虫对照品种sGK321,与抗虫对照R19抗虫性相当.遗传分析表明,转Bt Sck双价基因抗虫棉的抗性基因符合一对显性基因的分离规律.对转Bt Sck双价基因抗虫棉与R19及sGK321的杂交F1进行了抗虫性测定,所有的F1植株都表现出与转Bt Sck基因纯系亲本一致的抗虫性.转Bt Sck双价基因抗虫棉纯系312-5T2和332-2T2等位性测验证明:312-5T2与R19、sGK321的抗虫基因整合位点不连锁,表现为15∶1的自由组合比例;而332-2T2与R19中抗虫基因的整合位点表现连锁,与sGK321的抗虫基因表现自由组合.这为培育新的双价抗虫棉品种(系)提供了优良的育种材料. 相似文献
4.
Bt抗虫棉新品系毒蛋白表达差异分析 总被引：1，自引：1，他引：0
【目的】本研究旨在探讨部分抗虫棉不抗虫的原因。【方法】通过室内生物测定、卡那霉素、Bt-Cry1Ab/Ac试纸条、PCR分析、Southern blot检测和Bt蛋白含量检测等方法分析了23个抗虫棉新品系的抗虫性。【结果】生测结果表明供试品系间幼虫死亡率差异显著。卡那霉素、Bt-Cry1Ab/Ac试纸条、PCR分析和Southern blot检测结果表明,Bt基因在供试材料中整合并稳定遗传且为单拷贝。RT-PCR结果表明,各试验品系Bt基因相对表达量差异较大,花期叶片Bt基因的相对表达量最高;Bt蛋白检测结果发现抗虫棉Bt蛋白表达量在生育前期大于后期,营养器官中的大于生殖器官,不同年份Bt蛋白含量差异显著。幼虫死亡率与Bt基因相对表达量的相关系数仅为0.1745,与Bt蛋白的相关系数为0.7130,表明Bt蛋白含量越高,抗虫性越好;各组织器官的Bt蛋白与Bt基因的相对表达量相关系数为-0.3659~0.2542,二者表达趋势不一致,表明遗传背景对抗虫棉Bt蛋白的表达具有一定影响。且Bt基因可能在转录后受到调控导致Bt蛋白含量发生变化,从而影响了抗虫棉的抗虫性。【结论】这些结果有望为抗虫棉育种提供参考。 相似文献
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Bt基因在不同陆地棉基因型的表达研究 总被引：2，自引：0，他引：2
研究了Bt基因在陆地棉不同遗传背景下的表达差异。结果表明,(1)Bt基因在不同陆地棉基因型中均能充分表达,与常规(非抗虫)棉相比抗虫性极为显著。(2)不同陆地棉基因型品系(种)之间抗虫性比较,有差异且部分品系间达极显著水平。(3)Bt基因在不同陆地棉基因型的表达,前期(对二代棉铃虫的抗性)差异较大;中期(对三代棉铃虫的抗性)差异变小;后期(对四代棉铃虫的抗性)没有显著差异。(4)转Bt基因抗虫棉Bt基因的表达随棉花的生育进程呈逐渐降低趋势,即对二代棉铃虫的抗性>对三代棉铃虫的抗性>对四代棉铃虫的抗性。 相似文献
7.
采用本研究室建立的根癌农杆菌介导的高效遗传转化体系,转化苎麻优良品种芦竹青,获得了转Bt基因苎麻候选植株。通过PCR和Southern杂交等分子检测,证明Bt基因已经整合到部分候选植株基因组中。选取PCR和Southern杂交均为阳性的部分株系(T0)种植在大田,对这些植株进行室内抗虫鉴定并考察其主要农艺性状和品质性状,次年对T1代植株进行PCR和Southern杂交检测。结果表明, 与对照相比,T0代转Bt基因苎麻植株的抗虫性均强于对照,部分株系的抗虫性显著强于对照植株;且基本保持了亲本的优良性状;T1代植株中也含有Bt基因,表明Bt基因能稳定遗传,且T1代植株在大田的抗虫性明显强于非转基因植株。 相似文献
8.
抗虫杂交棉F1代与亲本Bt蛋白表达量及抗虫差异性研究 总被引：1，自引：0，他引：1
对转 Bt基因抗虫杂交棉正、反交 F1代与抗虫亲本的 Bt蛋白表达量及抗棉铃虫差异性研究表明 :抗虫亲本与其杂种 F1代均高抗棉铃虫 ,但抗虫亲本的抗虫性略好于其杂种 F1代 ,并且明显地高于非抗虫亲本 ;正、反交杂种 F1代间的抗虫性几乎没有差异。生长前期的抗虫性好于后期 ,同一时期嫩叶或侧枝生长点的抗虫性好于幼蕾。抗虫亲本叶片和花瓣的 Bt蛋白含量明显地高于其杂种 F1代 ,抗虫亲本功能叶的 Bt蛋白含量明显地高于其上部非功能叶 ,而杂种 F1代功能叶的 Bt蛋白含量则明显地低于其上部非功能叶。盛花期后至吐絮期前 ,叶片和花瓣的 Bt蛋白表达量明显增加 ,在抗虫亲本中表现最为明显。与叶片相比 ,在花瓣中检测到的 Bt蛋白含量极低。正、反交 F1代间的 Bt蛋白表达量差异较小或无规律可循 相似文献
9.
转基因抗虫棉Bt 毒蛋白表达量的传递方式研究 总被引：1，自引：0，他引：1
从卡那霉素抗性鉴定、抗虫性鉴定和 Bt毒蛋白含量测定等三个方面对转基因抗虫棉 Bt毒蛋白表达量的传递方式进行了研究。结果表明 ,转基因抗虫棉美棉 3 3 B和 GK-12对棉铃虫具有显著的抗性。盛蕾期饲喂美棉 3 3 B、GK-12棉株顶端叶片 72 h后 ,初孵棉铃虫幼虫死亡率分别为86.8%、75 .1% ,对照 TM-1、泗棉 3号、苏棉 12三个常规棉品种 (系 )初孵幼虫死亡率分别为10 .9%、13 .9%、9.2 %。美棉 3 3 B、GK-12盛蕾期功能叶片 Bt毒蛋白含量分别为每克鲜重 83 6.68ng、682 .5 6ng。饲喂美棉 3 3 B、GK-12与常规棉品种 (系 )杂种一代棉株顶端叶片 72 h后 ,初孵棉铃虫幼虫平均死亡率分别为 84.1%、77.2 % ,两个转基因抗虫棉品种与常规棉品种 (系 )杂种一代功能叶片的 Bt毒蛋白含量平均值分别为每克鲜重 82 0 .5 8ng、683 .77ng。转基因抗虫棉与常规棉杂种一代的抗虫性表现及 Bt毒蛋白表达量与转基因抗虫棉亲本非常接近 ,杂种二代群体 Bt毒蛋白检测阳、阴性反应植株的分离比例符合 3∶ 1,回交世代 BC1 群体 Bt毒蛋白检测阳、阴性反应植株的分离比例符合 1∶ 1,与抗虫性鉴定结果高度一致。转 Bt基因抗虫性状的遗传是受一对完全显性基因控制的 ,Bt基因与 NPT II基因是紧密连锁或完全连锁的。Bt毒蛋白表达量按照一对显 相似文献
10.
为了给转Bt基因棉花在科研和生产上多世代可持续利用提供依据,以转Bt基因棉花品种银山8号为试验材料,从2005年开始,连续16年进行跟踪研究。结果表明,随种植世代的增加,生物测定银山8号第2～4代棉铃虫幼虫校正死亡率、铃期叶片和铃期小铃杀虫蛋白表达量均有不同程度的线性上升趋势,同时皮棉产量线性回归也呈增长趋势;苗期叶片、蕾期叶片和蕾期小蕾杀虫蛋白表达量呈线性下降趋势。相关性分析表明,抗虫棉的抗虫性与皮棉产量间均呈正相关,株铃数和衣分是构成皮棉产量的重要因子,与皮棉产量的相关性分别达到极显著和显著水平。外源Bt基因转入棉花后能稳定遗传给后代,通过卡那霉素鉴定和系统选择可以持续保持转Bt基因棉花多世代的抗虫性,甚至通过提纯复壮逐年优选,可实现转Bt基因抗虫棉特定生育时期或部分棉花器官抗虫性的提高。 相似文献
11.
Bt基因转化玉米培育抗玉米螟自交系 总被引：7，自引：0，他引：7
采用基因枪法将苏云金杆菌(简称Bt)的杀虫毒蛋白基因(CryIA)导入东北春玉米自交系铁7922的幼胚中,诱导愈伤组织.抗虫基因的表达载体是pBI121,包含Bt杀虫毒蛋白基因和新霉素磷酸转移酶(NPTⅡ)基因,筛选标记bar由CaMV 35S启动子驱动.通过对后代植株的除草剂草丁膦(PPT)筛选、分子鉴定和ELISA法检测,证明外源基因已经整合到玉米基因组中并可以稳定遗传.对转基因株系进行田间接玉米螟虫卵,调查不同生长时期的危害指数,最终筛选出一批抗玉米螟自交系. 相似文献
12.
转Bt基因玉米对亚洲玉米螟的抗性评价 总被引：7，自引：0，他引：7
转Bt基因抗虫玉米为诺华公司含Bt11转化系的杂交种NX4777,能全株表达Cry1Ab杀虫蛋白,以其非转基因受体玉米品种NX4906为对照。在玉米生长至心叶中期(50~60cm高)、抽雄前期和抽丝散粉期分别进行人工接亚洲玉米螟[Ostrinia furnacalis (Guenée)]初孵幼虫40~60头。以食叶级别、存活幼虫数、蛀孔数、隧道长度和雌穗被害级别等,分别作为评价玉米心叶期和穗期的抗性标准。结果表明,Bt玉米食叶级别仅为1级,显著低于对照的7级。Bt玉米平均每株存活幼虫0.04~0.20头、蛀孔0.11~0.15个、隧道长度0.13~0.41cm,雌穗被害级别为0,植株折茎率为0,而对照玉米平均每株存活幼虫6.19~12.41头、蛀孔4.48~7.05个、隧道长度12.41~24.09cm,雌穗被害级别为5.9,植株折茎率高达73.6%~95.5%。室内生测结果亦表明,取食Bt玉米心叶、雄穗的初孵幼虫3d全部死亡,取食花丝7d死亡率达到99%,相比之下取食常规玉米的存活率在88.7%以上。表明表达Cry1Ab杀虫蛋白的Bt对亚洲玉米螟具有很高的抗虫性,能够保护玉米全生育期免受玉米螟的危害。 相似文献
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The European corn borer (ECB) has become a serious pest for maize cultivation in Central and Western Europe. This study was
conducted to evaluate resistance against ECB in a cross section of the European maize germplasm. A total of 115 flint and
dent inbreds were evaluated for resistance under protected and artificial infestation conditions at two German sites in 1993.
Resistance to ECB was assessed by damage rating before harvest, tunnel length in dissected stalks, and relative yield under
infestation compared to protected control plots. Means for damage rating of stalks were significantly higher in flint lines
than in dent lines. Artificial infestation reduced grain yield by 31% in flint lines and 20% in dent lines. Significant genotypic
variances among lines and high genetic ratios (GR) were found in both flint and dent lines for all agronomic and ECB resistance
traits. Exceptions were relative yield of stover in both germplasm groups and tunnel length in flint lines. Phenotypic correlations
between agronomic and resistance traits were moderate to low. Damage rating was negatively correlated with days to silking
and dry matter yield of stover in both germplasm groups. In vitro digestibility of stover was not associated with ECB resistance.
Correlations of relative grain yield with tunnel length below the ear and damage rating were significantly negative, even
though their magnitude was low. Our results indicate substantial genetic variation among European inbred lines for resistance
to the univoltine ECB. Among all resistance traits measured, damage rating of stalks is best suited for assessment of ECB
resistance in breeding programs because of its easy recording and high genetic ratio.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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人工模拟“庇护所”和“非庇护所”条件下棉铃虫对Bt棉的抗性演变 总被引：1，自引：1，他引：0
人工饲料中添加转Bt基因杂交棉品种中棉所29棉仁粉,建立杂交和自交品系分别模拟“庇护所”和“非庇护所”条件,连续多代进行棉铃虫对Bt棉的抗性筛选。结果表明：经11代筛选,棉铃虫自交品系和杂交品系的幼虫死亡率下降,体重和体长增加,幼虫历期缩短,化蛹率和羽化率增加。依据幼虫死亡率、体重、体长、幼虫历期以及化蛹率、羽化率等生命参数的演变,相较杂交品系,自交品系对Bt棉能更快地产生适应性。11代筛选后,利用不同抗性品系棉铃虫进行转基因棉抗虫性评估,结果表明：相比敏感品系,杂交品系测定4个转基因棉品种的抗虫性等级未发生变化,而中棉所29等3个转基因棉品种对棉铃虫自交品系的抗性下降了1个等级。对Bt制剂的抗性测定结果表明,杂交品系的抗性倍数为2.5215,自交品系的抗性倍数为9.3876。据本研究初步结果,利用“庇护所”策略延缓棉铃虫对转Bt基因棉的抗性产生是可行的。 相似文献
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There is not any conclusive result about the most suitable trait for performing pedigree selection for improving maize performance against corn borer attack: tunnel length or stalk breakage. We have used simultaneously both selection traits in the same genetic backgrounds and the objective of the present work has been to compare the suitability of pedigree selection for tunnel length and stalk breakage in order to obtain inbred lines that per se and in hybrid combination show lower stem damage and good agronomic performance. Tunnel length is better as selection criteria for increasing resistance to corn borers, but, in some genetic backgrounds, genetic linkage between yield and stalk tunneling by stem borers could make that selection for reduced tunnel length resulted in significant yield reductions. 相似文献
16.
将组成型表达的玉米泛素启动子与苏云金杆菌杀虫蛋白基因Bt连接，插入根瘤农杆菌双T-DNA质粒，构建一个T-DNA结构域含有抗潮霉素选择标记基因hyg，另一个T-DNA结构域含有抗虫基因的双T-DNA单子叶植物表达载体，用以转化农杆菌菌株，再通过共培养转化玉米胚性愈伤组织。通过潮霉素培养基抗性筛选，用特异PCR扩增和Southern杂交检测，从分化再生的T0代植株中，鉴定出4个转化Bt基因的阳性植株。目前，正结合进行田间分离纯合和DNA分子鉴定，培育去除选择标记基因的转基因抗虫玉米自交系。 相似文献
17.
The univoltine European corn borer (ECB) has become a major limiting factor for maize (Zea mays L.) production in central Europe. The objective of this study was to survey the genetic variation for ECB resistance in European elite maize germplasm. Eighteen flint and 23 dent inbreds were screened under artificial ECB infestation at two locations in 1993 and 1994. Resistance was assessed by damage rating of broken plants, measurement of tunnel length in dissected stalks, and yield reduction in infested plots relative to insecticide-protected control plots. Flint lines showed significantly greater means for damage rating than dent lines with grain yield reduction of 35% and 24%, respectively. Significant genotypic variances among lines and high heritabilities were found for agronomic traits and damage rating. Heritabilities were intermediate for tunnel length and relative grain yield. Significant associations of days to silking, ear dry matter content, and dry matter yield of the whole plant with damage rating and tunnel length suggested a better resistance in late-maturing, high-yielding inbreds. Genotypic correlations of relative grain yield with tunnel length and damage rating ranged between ?0.46 and ?0.72. Partial correlations, eliminating the effect of flowering time, confirmed these associations. Damage rating of stalks is the most suitable trait for evaluation of ECB damage owing to its high heritability and easy recording. Tunnel length below the primary ear is a useful trait for assessing antibiosis because it is not correlated with days to silking. Inbreds with extreme resistance and susceptibility were identified which can be used as parents for establishing breeding and QTL mapping populations. 相似文献
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19.
基因枪法转Bt/GNA基因超甜玉米植株的获得及抗虫性鉴定 总被引：1，自引：0，他引：1
以超甜玉米自交系S1幼胚诱导的愈伤组织为受体材料,利用基因枪法共转化Bt和GNA基因,获得了128株再生植株,移栽成活31株,其中26株可育.经PCR检测外源基因整合进玉米基因组中,含Bt基因11株,其中有5株能扩增出GNA基因.Southern杂交证明,目的基因已经整合在玉米基因组中.对9株转基因T1代植株叶片室内接虫抗性鉴定,结果显示,5株转基因植株叶片饲喂6 d后明显抑制玉米螟幼虫的发育,幼虫体重增加较少,幼虫死亡率高,表现较强的抗性. 相似文献
20.
The spotted stem borer, Chilo partellus, is one of the most important pests of sorghum, and host plant resistance is an important component for the management of
this pest. Most of the sorghum hybrids currently under cultivation are based on cytoplasmic male-sterility (CMS). In order
to develop a strategy for resistance to stem borer, we studied the traits associated with resistance, and their nature of
gene action in F1 hybrids derived from resistant, moderately resistant, and susceptible CMS and restorer lines. The hybrids based on stem borer-resistant,
moderately resistant, or susceptible CMS and restorer lines were equally resistant or susceptible as the parents for leaf
feeding [Damage rating (DR) 5.8 to 6.6 vs. 5.9 to 6.6], and had significant and decreasing trend in deadheart formation (resistant
CMS × resistant restorer lines < moderately resistant CMS × moderately resistant restorer lines < susceptible CMS × susceptible
restorer lines), respectively. Proportional contributions of restorer lines were greater than those of the CMS lines for leaf
feeding, deadhearts, recovery and overall resistance, stalk length, nodes per plant, stem borer holes per plant, and peduncle
tunneling. The general (GCA) and specific combining ability (SCA) estimates suggested that leaf feeding score, number of nodes,
overall resistance score, panicle initiation, recovery score, and stalk length (dominance type of gene action) have been found
to be associated with resistance to spotted stem borer, governed by additive type of gene action, their correlation and direct
effects in the same direction, and explained 65.3% of the variation in deadhearts, and thus could be used as marker traits
to select and breed for resistance to C. partellus in sorghum. The parents having significant SCA effects for two or more resistance traits for either or more parents have
also been discussed for their use in the stem borer resistance breeding. 相似文献