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1.
以13个基因型小麦的幼胚愈伤和新春9号幼胚为受体材料,利用农杆菌菌株C58C1(含有GUS外源基因)进行侵染,通过组织化学染色的方法统计GUS基因瞬时表达率,以筛选对农杆菌敏感的小麦基因型及适宜的菌液浓度和侵染时间,优化农杆菌转化的条件,为逐步建立小麦农杆菌的转化体系奠定基础.结果显示,不同基因型小麦GUS瞬时表达率在0～95.9％之间,其中新春9号、小偃328和济麦19的GUS基因瞬时表达率均在90％以上;对新鲜幼胚和预培养7d的幼胚愈伤组织分别侵染30 min和60 min,前者GUS阳性率均为0,后者分别为92.0％和95.9％,说明预培养可以显著提高受体材料对农杆菌的敏感性,提高转化效率;在不同菌液浓度(OD600=0.5、1.0、1.5)侵染时,GUS基因瞬时表达率分别为93.4％、94.0％和87.6％,不同侵染时间(15,30,60 min)下,GUS基因瞬时表达率分别为94.6％、92.0％和95.9％.综合考虑认为,适宜的小麦农杆菌转化条件为幼胚经过4～7 d预培养,在菌液浓度OD600=1.0时进行15 min的侵染. 相似文献
2.
农杆菌介导玉米遗传转化体系的优化 总被引：3，自引：0，他引：3
以自交系18H、M017、9046和178的幼胚为材料,进行愈伤组织诱导,用根癌农杆菌EHA105和LBA4404对获得的Ⅱ型胚性愈伤组织进行转化,导入TERFI-LeERF2基因,并对农杆菌介导玉米遗传转化体系的条件进行了优化。结果表明：使用毒性较强的根癌农杆菌EHA105侵染愈伤组织;农杆菌浓度在0D600=0．6,侵染时间20min;侵染后的愈伤组织经7d的恢复培养后,先进行低浓度选择压的筛选培养,再进行高浓度选择压的筛选培养,以及在筛选培养基中添加10mg／LAgNO,提高了抗性愈伤组织的获得率。获得植株经PCR鉴定表明,外源基因已经整合到玉米自交系18H的基因组中。 相似文献
3.
针对转基因小麦成熟胚转化体系转化效率低,从提高成熟胚愈伤组织质量入手,调节小麦的基因型、成熟胚的培养时间、菌液浓度、侵染时间以及愈伤组织大小几个因素,提高成熟胚转化体系的转化效率;通过农杆菌介导,将目的基因转入到受体小麦成熟胚诱导的愈伤中,利用gus检测的方法检验转化效率;继代360d、自然生长5mm大小的受体小麦品种扬麦10,在菌液浓度OD600为0.8,侵染40min下,gus基因表达率达50%左右,接近幼胚的gus基因表达率。通过对体系的几个因素的调节,提高了农杆菌介导成熟胚转化体系的效率。 相似文献
4.
为了丰富栽培小麦转化受体基因型,优化农杆菌转化小麦幼胚的技术体系,本研究评价了21份四川优良小麦品种(系)的幼胚再生能力,并以筛选到的最佳受体为材料,利用正交设计研究了预培养时间、菌液浓度、侵染时间、抑菌剂浓度、共培养方式和农杆菌菌株等易互作的6个因素对幼胚抗性愈伤再生率的影响,并探讨了筛选剂卡那霉素(Kanamycin)的适宜浓度。结果表明,21份材料中‘5157’、‘川农16’、‘R364’、‘川麦42’‘、内麦8号’的绿苗率超过了30%,可作为优良转基因受体加以利用。以‘5157’为受体材料建立农杆菌转化体系,正交试验表明菌液浓度和农杆菌菌株对幼胚抗性愈伤再生率的影响达到显著水平,而其他因素影响不显著。经优化后的遗传转化体系为:农杆菌菌株用C58C1,幼胚不经预培养,在菌液OD值为0.4的农杆菌中侵染40 min,共培养介质选用培养基,抑菌剂为200 mg/L羧卞青霉素(Carbenicillin),适宜的卡那霉素筛选浓度为50 mg/L。本研究为四川小麦遗传转化提供了5个候选基因型,为建立和优化小麦幼胚转化技术体系提供了理论依据,同时初步建立了农杆菌转化小麦幼胚的技术体系。 相似文献
5.
以籼稻品种中籼3037为材料,研究了影响农杆菌介导转化籼稻的几个重要因素,结果表明：MSⅡ培养基作为愈伤组织的诱导培养基好于N6D2;以幼胚为外植体,愈伤组织的诱导率和转化率明显比成熟胚和幼穗高;根癌农杆菌菌株EHA105介导的基因转化率高于AGLO;当以幼胚为外植体,根癌农杆菌菌株E-HA105介导的中籼3037的转化率为18.0%,而AGLO为5.5%;EHA105菌液OD值为0.6,浸染时间14min为中籼3037合适的转化参数;在分化培养基MSR中添加20g/L的山梨醇有利于抗性愈伤组织的分化。在此基础上,采用优化的农杆菌介导法转化中籼3037及其来源的突变体A和B,获得了转化植株,PCR、Southern杂交和T1代遗传分析表明外源基因已经整合到水稻基因组中。 相似文献
6.
农杆菌介导玉米遗传转化体系的研究 总被引：4，自引：1，他引：3
为了提高玉米自交系的遗传转化率,建立农杆菌介导玉米遗传转化体系。通过农杆菌菌株EHA105侵染玉米自交系7922的胚性愈伤组织,以β-葡萄糖苷酶基因(GUS)为报告基因研究几种因素对遗传转化体系的影响。结果表明：（1）当农杆菌浓度OD600=0.6时,GUS瞬时表达率最高为35.3%;（2）当侵染时间为15 min时,GUS瞬时表达率最高为37.3%;（3）当共培养时间为3天和恢复培养时间为4天时,GUS瞬时表达率最高为29.1%。因此选择农杆菌浓度OD600=0.6,侵染时间15 min,共培养3天和恢复培养4天,为最佳遗传转化条件。通过优化影响遗传转化体系的因素,初步建立了农杆菌介导的玉米遗传转化体系。 相似文献
7.
农杆菌介导小麦遗传转化条件的优化 总被引：2，自引：1，他引：1
本研究以普通小麦品种扬麦15和Alondra's的幼胚产生的愈伤组织为受体材料,通过检测gus基因的瞬时表达情况,研究了农杆菌介导的主要影响因子的转化效率。研究结果表明,在相同条件下两个品种农杆菌侵染的最佳条件不尽相同。扬麦15的最佳优化条件为:先在不添加AS的培养基中预培养,然后在菌液浓度为OD600=0.2、AS浓度为100μmol/L条件下侵染10min,最后在25℃共培养1d;Alondra's则在菌液浓度为OD600=0.5、共培养时间为2d时,表现出最佳的gus基因瞬时表达率。为小麦的遗传转化提供参考。 相似文献
8.
适用于4种玉米基因型的农杆菌转化方法的探讨 总被引：1，自引：0，他引：1
利用改良的农杆菌培养液、侵染液、共培养培养基和筛选培养基等技术体系,对4个玉米基因型Hi-Ⅱ、H99、国内2个优良自交系R18-599和齐319的新鲜幼胚、预培养幼胚、胚性愈伤组织进行了农杆菌转化研究,并比较了不同共培养方式对农杆菌侵染不同玉米组织的影响。结果表明,Hi-Ⅱ新鲜幼胚在固体培养基上共培养和滤纸上共培养后GUS基因瞬间表达率分别为83.7%和4.4%,前法优于后法;Hi-Ⅱ新鲜幼胚、预培养3 d的幼胚愈伤组织经农杆菌侵染后GUS基因瞬间表达率分别为83.7%和12.1%,新鲜幼胚比预培养的幼胚愈伤组织更适合于农杆菌转化。用上述优化的条件对另外3个不同基因型的2种不同外植体H99、齐319(新鲜幼胚)和R18-599（胚性愈伤组织）进行遗传转化,共培养3 d后的GUS基因瞬间表达率分别为65.2%、52.6%和58%,表明该转化体系适合于上述4种基因型的幼胚和胚性愈伤组织的遗传转化。此外农杆菌侵染Hi-Ⅱ新鲜幼胚后经在含巴龙霉素25~100 mg L-1培养基上3轮选择后,抗性愈伤组织获得率为2.4%,近似反映了本研究的遗传转化率。其他3个基因型的抗性愈伤也正在筛选中。结果初步表明,本研究建立的农杆菌介导的玉米遗传转化体系可能对4种基因型均适用。 相似文献
9.
10.
小麦TaPHR1基因表达载体的构建 总被引：2，自引：0，他引：2
本研究以植物表达载体pROK2-Ubi为基础,设计带有酶切位点KpnⅠ和BamHⅠ的一对引物,从载体pAC25上扩增到目的基因TaPHR1。酶切回收后与同样双酶切的表达载体pROK2-Ubi连接,获得新的表达载体pTaPHR1,并将所构建的载体导入根瘤农杆菌LB4404菌株。另外以小麦品种济麦22为受体,利用农杆菌介导小麦成熟胚愈伤组织的遗传转化体系和构建的表达载体进行了初步的遗传转化。实验结果表明选择抗生素选择压Kan50mg/L,接种菌液浓度为OD600=0.6,侵染时间为60min时,抗性愈伤的获得率最高,达到4.08%,抗性愈伤组织的分化率达到3.28%。本研究为农杆菌介导缺磷响应基因TaPHR1的小麦遗传转化和小麦磷高效遗传改良研究提供了研究数据。 相似文献
11.
Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts 总被引：7，自引：0，他引：7
Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed. 相似文献
12.
C. J. Spurr D. A. Fulton P. H. Brown R. J. Clark 《Journal of Agronomy and Crop Science》2002,188(4):275-280
Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality. 相似文献
13.
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
14.
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
15.
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
16.
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
17.
Summary Reactions of 13 grasses to Meloidogyne naasi varied with species; ryegrasses, fescues and their hybrids were generally susceptible and cocksfoot and timothy resistant. Marked variation in host resistance levels occurred between genotypes within cultivars.Selection of single plants, followed by tests on replicate tillers, identified resistant and susceptible genotypes in both Italian and perennial ryegrass cultivars. Resistant plants had few nematode-induced galls and fewer females and eggs than susceptibles. There was more or less continuous variation, with many genotypes intermediate between extremes of resistance and susceptibility. Selected resistant and susceptible genotypes are of use in assessing variation in nematode populations and as controls for breeding and selection programmes. 相似文献
18.
W. Erskine M. Tufail A. Russell M. C. Tyagi M. M. Rahman M. C. Saxena 《Euphytica》1993,73(1-2):127-135
Lentil production is limited by lack of moisture and unfavorable temperatures throughout its distribution. Waterlogging and salinity are only locally important. Progress has been made in breeding for tolerance to drought through selection for an appropriate phenology and increased water use efficiency and in breeding for winter hardiness through selection for cold tolerance.The diseases rust, vascular wilt, and Ascochyta blight, caused by Uromyces viciae-fabae, Fusarium oxysporum f. sp. lentis, and Ascochyta fabae f. sp. lentis, respectively, are the key fungal pathogens of lentil. Cultivars with resistance to rust and Ascochyta blight have been released in several countries and resistant sources to vascular wilt are being exploited. Sources of resistance to several other fungal and viral diseases of regional importance are known. In contrast, although the pea leaf weevil (Sitona spp.) and the parasitic weed broomrape (Orobanche spp.), and to a lesser extent the cyst nematode (Heterodera ciceri), are significant yield reducers of lentil, no sources of resistance to these biotic stresses have been found. Directions for future research in lentil on both biotic and abiotic stresses are discussed. 相似文献
19.
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
20.
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