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1.
《分子植物育种》2016,(4)
制约大豆生产的因素很多,土壤盐渍化就是其中之一。大量研究证明过表达NHX逆向转运蛋白可以提高植物的耐盐性。为获得耐盐性良好的转基因大豆材料,我们将大豆Na~+/H~+逆向转运蛋白(GmNHX1)基因构建到植物表达载体pCAMBIA3300上,应用农杆菌介导法将GmNHX1导入大豆品种黑农56和黑农59中,共获得18个转基因株系,并对T_1代转基因株系进行了PCR和实时荧光定量PCR检测。PCR结果表明转基因后代株系呈阳性的植株有4株;经Real-time PCR检测该4个PCR阳性转基因株系的G NHX1基因表达水平均高于对照株系;200 mmol/L NaCl溶液的盐试结果表明;对照植株的生长速度滞缓于4个转基因株系的生长速度。 相似文献
2.
pCB-zeolin-GFP表达载体的构建及瞬时表达 总被引：1，自引：1，他引：0
为利用荧光蛋白基因GFP检测外源基因在转基因植株中的表达和定位,构建含有GFP基因的植物表达载体pCB-zeolin-GFP。在目的基因的开放阅读框（ORF）两端设计引物,并引入酶切位点和保护碱基,用PCR方法从pDHA扩增得到zeolin基因的全长,克隆到中间载体pMD18-T,分别用NcoⅠ和BglⅡ2种限制性内切酶酶切重组质粒和经过改良的pCAMBIAI1302植物表达载体,经回收、连接、转化、鉴定后,利用基因枪转化法将重组载体转入洋葱表皮细胞,通过共聚焦显微镜检测绿色荧光蛋白在洋葱表皮细胞中的瞬时表达。构建了zeolin基因与绿色荧光蛋白（GFP）融合的植物表达载体pCB-zeolin-GFP,并在洋葱中得到了表达。构建的融合植物表达载体pCB-zeolin-GFP正确,该载体的成功构建为今后进行基因转移、基因功能研究及培育新品种奠定了基础。 相似文献
3.
适于植物转基因体系优化的绿色荧光蛋白表达载体构建及其瞬时表达验证 总被引：1，自引：1，他引：0
在建立和优化植物的转基因体系时,通常期望转化载体同时具有适用植株范围广、尺寸较小、具有多重选择标记或报告基因等优点。为此,通过Gateway技术,构建了含有绿色荧光蛋白基因GFP的植物表达载体pGWB14-GFP,并经PCR鉴定和测序验证了序列的准确性。利用基因枪转化法,将该载体转入洋葱表皮和小麦叶片中,通过荧光显微镜检测,发现绿色荧光蛋白在洋葱表皮细胞和小麦叶片表皮细胞中均能瞬时表达,表明pGWB14-GFP在双子叶和单子叶植物中都能正常发挥功能。本载体在双子叶和单子叶植物植物中都可使用,大小低于17 kb,含有GFP报告基因,具有两套卡那霉素和潮霉素选择标记基因,用于在细菌和植物中筛选,为植物转基因体系的构建和优化提供了功能强大的转化载体。 相似文献
4.
水稻稻瘟病是由子囊菌(Magnaporthe oryzae)引起的水稻灾害性病害。培育抗病品种是防治稻瘟病最经济有效的措施之一。水稻Pi9抗稻瘟病基因来源于小粒野生稻并已被克隆和应用于转基因育种。为了提高无选择标记转基因植株的选择效率,将绿色荧光蛋白(GFP)用作可视遗传标记,对双菌株共转化系统进行改良:目的基因载体携带Pi9抗稻瘟病基因;标记基因载体用潮霉素磷酸转移酶(HPT)作为植物转化选择标记,用GFP作为负选择标记,筛除标记基因分离植株。两种载体的农杆菌转化株混合,分别与水稻品种‘浙恢414’、‘浙粳22’、‘浙11B’、‘日本晴’、‘空育131’和‘粤泰B’的愈伤组织共培养,然后从5%~38.3%的起始愈伤组织筛选获得了转化愈伤组织(HPT+GFP+)。对T0植株进行Pi9基因PCR检测,11.8%~77.8%的T0植株为共转化植株(HPT+GFP+Pi9+)。对共转化植株T1代进行绿色荧光检测,筛选阴性植株(GFP-),再通过PCR筛选Pi9+植株。根据13个T1群体的研究结果,61%的共转化植株在T1代分离出无选择标记转基因植株(HPT-GFP-Pi9+)。转Pi9的无选择标记植株和后代株系对水稻稻瘟病呈抗病反应。因此,本研究通过GFP标记提高了双菌株共转化系统的选择效率,转Pi9的无选择标记水稻株系为水稻抗稻瘟病育种提供了有用的遗传资源。 相似文献
5.
以甘露糖作为筛选剂的棉花遗传转化 总被引：14，自引：0，他引：14
以pmi基因作为筛选标记基因,以甘露糖作为筛选剂,通过农杆菌介导法将GFP基因导入棉花细胞并得到再生植株,经过PCR检测、Southern杂交证实外源基因已经整合到棉花基因组中,Westbloting与荧光显微镜检测证明GFP基因得到表达。本文研究讨论了甘露糖作为棉花转化细胞的筛选剂在农杆菌介导的转化中的应用浓度及方法,即:甘露糖的筛选浓度在30～50g·L 1之间,在愈伤组织诱导初期适当低一点,随着愈伤组织的生长而加大筛选浓度。由于甘露糖不利于再生胚的分化,当愈伤组织转入分化培养基时,要以葡萄糖代替甘露糖。 相似文献
6.
擎天凤梨的愈伤组织获取较为困难、而且愈伤分化缓慢且极易死亡,使用农杆菌介导转基因面临着受体难以获得、侵染后愈伤死亡率高、转化时间长以及成本高等问题。使用基因枪介导的转基因方法相对来说更为直接快速,但也面临转化效率不高、插入基因表达定位不清等问题。本研究以擎天凤梨组培苗的茎尖生长组织为材料,以基因枪介导结合GFP荧光蛋白基因,对转化中的主要影响因子进行研究,建立并优化了遗传转化体系,获得了转化再生植株,并利用GFP荧光蛋白基因的荧光特性,分析确定了外源基因的表达定位,为进一步利用分子育种手段培育擎天凤梨新品种打下基础。 相似文献
7.
8.
根癌农杆菌介导苎麻转绿色荧光蛋白（GFP）基因植株再生 总被引：2，自引：0，他引：2
以苎麻优良品系“5041-3”子叶作为受体,利用根癌农杆菌介导法进行了绿色荧光蛋白基因的遗传转化研究,农杆菌菌株为EHA105,重组质粒为pBIN m-GFP5-ER。经农杆菌浸染后的子叶外植体经过共培养、选择培养、伸长培养和生根培养后再生出抗性植株,对抗性植株的再生过程进行了GFP荧光检测,结果表明,GFP基因能在苎麻中强烈表达,证明GFP基因能够在苎麻遗传转化中得到应用。对抗性植株的Southern杂交检测证实外源基因已整合到苎麻基因组中。 相似文献
9.
《分子植物育种》2017,(5)
甘蔗是重要的糖料作物,也是重要的能源作物。由于复杂的遗传背景、较低的可育性和缺乏优异种质资源等因素给甘蔗品种改良带来了很大的困难,通过转基因技术为甘蔗品种改良提供了有效的途径。而转基因植物的遗传稳定性是植物转基因育种过程中一项重要的研究课题。本研究通过一次多基因遗传转化,同时将四个目的基因导入甘蔗品种新台糖22号中。通过PCR检测,对转基因甘蔗T_0、T_1和T_2代的遗传稳定性进行分析;通过外源蛋白快速试纸条检测,对转基因植株四个外源基因中的基因(bar抗除草剂基因和cry1Ab抗虫基因)蛋白表达进行检测。证明一次四基因遗传转化的甘蔗转基因株系,在T_0代出现了部分基因丢失的现象,且T_0代甘蔗的主茎和分蘖可能为不同的转基因事件。但如果是四个基因均能完整整合到基因组中的株系,则四个基因可以在T_0、T_1和T_2代中得到稳定遗传,也可以稳定的检测得到外源基因蛋白的表达。 相似文献
10.
根癌农杆菌介导小麦成熟胚的遗传转化研究 总被引：2，自引：1，他引：1
以小麦成熟胚为转化受体,采用农杆菌介导的遗传转化技术,建立了小麦的转基因株系.结果表明,建立的小麦遗传转化和植株再生体系,在选择压下由侵染的小麦成熟胚中,获得的抗性愈伤组织频率为57.50%,愈伤组织分化频率为8.58%,植株分化比例为0.83%.对再生植株中报告基因Gus的PCR检测结果表明,供试3株小麦的PCR均为阳性.与未进行遗传转化的对照植株相比,再生植株的Gus活性明显提高.表明再生的供试植株均为转基因植株.因此,本研究建立的小麦成熟胚遗传转化和再生体系,为今后小麦的高频转基因系的获得提供了参考依据. 相似文献
11.
Jens Jensen 《Euphytica》1979,28(1):47-56
Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans. 相似文献
12.
G. H. Kroon 《Euphytica》1994,76(1-2):125-125
Summary
K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes. 相似文献
13.
Hongyan WEI Jian ZHOU Jinguang LU Jingzheng SONG Qiongxi YU Zhihong JIANG 《Medicinal Plant》2019,(6):27-29
[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol... 相似文献
14.
Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed. 相似文献
15.
D. A. Bond G. J. Jellis G. G. Rowland J. Le Guen L. D. Robertson S. A. Khalil L. Li-Juan 《Euphytica》1993,73(1-2):151-166
Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses. 相似文献
16.
Chunli TANG Fengxian ZHAO Hongxia CHEN Jiabao MA Jiangcun WEI Meiyan QIU Zhen XIE 《Medicinal Plant》2019,(6):60-65
[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination... 相似文献
17.
Mohammed Riyazaddin Polavarapu B. Kavi Kishor Are Ashok Kumar Belum V. S. Reddy Rajendra S. Munghate Hari C. Sharma 《Plant Breeding》2015,134(4):423-436
Sorghum shoot fly, Atherigona soccata, is one of the important pests of postrainy season sorghums. Of the 90 sorghum genotypes evaluated for resistance to this pest, RHRB 12, ICSV 713, 25026, 93046 and 25027, IS 33844‐5, Giddi Maldandi and RVRT 3 exhibited resistance in postrainy season, while ICSB 463, Phule Anuradha, RHRB 19, Parbhani Moti, ICSV 705, PS 35805, IS 5480, 5622, 17726, 18368 and 34722, RVRT 1, ICSR 93031 and Dagidi Solapur showed resistance in rainy season, suggesting season‐specific expression of resistance to A. soccata. ICSB 461, ICSB 463, Phule Yasodha, M 35‐1, ICSV 700, 711, 25010, 25019 and 93089, IS 18662, Phule Vasudha, IS 18551 and 33844‐5 and Barsizoot had fewer deadhearts than plants with eggs across seasons, suggesting antibiosis as one of the resistance mechanism. Five genotypes exhibited resistance with high grain yield across seasons. Correlation, path and stepwise regression analyses indicated that leaf glossiness, seedling vigour, trichome density, oviposition and leaf sheath pigmentation were associated with the expression of resistance/susceptibility to shoot fly, and these can be used as marker traits to select and develop shoot fly‐resistant sorghums. 相似文献
18.
E. Keep 《Euphytica》1986,35(3):843-855
Summary Cytoplasmic male sterility (cms) is described in the F1 hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf
1) and recessive for the other (Rf
2).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph
3(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf
1(one of two dominant additive genes controlling early season leafing out) indicated that Rf
1and Rf
2were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf
1, and 0.15 for Lf
1-Sph
3were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph
3.It is suggested that competitive disadvantage of lf
1-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph
3. Poor performance of lf
1- (and possibly lf
2-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off). 相似文献
19.
[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho... 相似文献
20.
Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed. 相似文献