3个紫花苜蓿乙烯应答因子基因的克隆与分析     

陈婷婷  杨青川  康俊梅  丁旺  张铁军  张新全 《中国农业科学》2012,45(4):815-822

【目的】克隆3个紫花苜蓿乙烯应答因子基因,分析其在不同条件下的表达特性;构建植物表达载体并转化烟草,对转基因烟草的耐盐性进行初步鉴定。【方法】根据已获得的cDNA序列设计引物,扩增MsERF5、MsERF8和MsERF11的DNA序列,并分析基因结构;利用半定量RT-PCR技术分析其组织表达特异性和胁迫条件下的表达特性;通过农杆菌介导的方法转化烟草,鉴定盐胁迫条件下转基因烟草的表型和生理生化特性,初步验证其功能。【结果】MsERF5、MsERF8和MsERF11都不包含内含子。MsERF5在根、叶和花蕾中的表达量高于茎和花;MsERF8在根和叶中的表达量高于茎、花蕾和花;MsERF11在叶中的表达量最高;3个基因都能被多种非生物胁迫（盐、干旱、铝）和激素（脱落酸、赤霉素、乙烯利、水杨酸和茉莉酸甲酯）诱导,但表达模式不同。在盐浓度为200 mmol•L-1的筛选培养基上只有导入目的基因的愈伤组织能产生不定芽,250 mmol•L-1 NaCl处理再生植株10 d,转基因植株和野生型植株叶片的电导率和可溶性糖含量均呈现上升趋势,但野生型植株叶片的电导率显著高于转基因植株,可溶性糖含量则显著低于转基因植株（P＜0.05）;转基因植株和野生型植株叶片的叶绿素含量均呈现下降趋势,野生型植株叶片叶绿素含量显著低于转基因植株（P＜0.05）;盐胁迫条件下,转化不同基因的再生植株的电导率、叶绿素和可溶性糖含量没有显著差异（P＞0.05）。【结论】3个紫花苜蓿乙烯应答因子基因都不包含内含子,其表达具有组织特异性,都能被多种非生物胁迫和激素诱导,能够使烟草愈伤组织在含盐培养基上形成不定芽并最终产生转基因植株,且转基因植株的耐盐性高于野生型植株。  相似文献   

紫花苜蓿白粉病病原鉴定   总被引：11，自引：2，他引：11  

李敏权  张自和  柴兆祥  李金花  骆惠生 《甘肃农业大学学报》2002,37(3):303-306,328

通过对甘肃定西县一年生紫花苜蓿白粉病蜡叶标本上的白粉菌的显微观察和计测,该菌闭囊壳直径85.5～128.3 m,平均直径95.7 m。每个子囊壳附属丝平均数量为27条,多数为20～30条。每个子囊壳含子囊6～8个,多数为6个,子囊大小为85.5m32.1 m。子囊内有子囊孢子2～4个,绝大多数为2个。167个子囊孢子计测结果,大小为32.1～21.4 m。附属丝褐色,多数基生,少数侧生,菌丝状,长度为子囊壳直径的0.6～1.6倍。经鉴定苜蓿上的白粉菌为蓼白粉菌(Erysiphe polygoni DC.)  相似文献   

抗臌胀病紫花苜蓿研究进展   总被引：10，自引：0，他引：10  

刘志鹏杨青川  呼天明 《四川草原》2003,(5):7-8

综述了紫花苜蓿中导致瘤胃臌胀病的臌胀因子,包括浓缩单宁、可溶性蛋白和皂角苷,同时阐述了抗臌胀病紫花苜蓿育种研究进展。  相似文献   

直立型扁蓿豆对干旱胁迫和复水的响应及适应策略     

乌日娜  石凤翎  徐舶 《中国生态农业学报》2020,28(12):1901-1912

以直立型扁蓿豆[Medicago ruthenica（L.）Sojak.cv.Zhilixing]为材料,于苗期连续干旱处理12 d后复水4 d,研究直立型扁蓿豆幼苗形态结构特征、生理代谢及生物量分配对干旱胁迫及复水的响应,揭示直立型扁蓿豆对干旱胁迫及复水的适应策略。结果表明：随着干旱胁迫时间延长,直立型扁蓿豆叶片气孔开放率逐渐降低,处理9 d后气孔及表皮细胞密度比正常浇水处理（CK）分别增加48.5%和36.6%,形成小而密的表皮细胞和气孔。生理上,除MDA含量随胁迫时间的延长逐渐增加外,其余指标均先升高后降低,干旱胁迫9 d时达最高,SOD、POD、叶绿素、可溶性糖、可溶性蛋白及脯氨酸分别较CK提高88.9%、111.2%、86.7%、140.5%、147.8%和124.6%。同时,生物量随胁迫时间的延长先增大后减小,于干旱胁迫9 d达最大值,比CK增加16.4%,总的分配格局表现出地上生物量投资高于地下,地下生物量投资比例随胁迫时间的延长逐渐增加,而地上生物量变化与其相反。复水后各指标均能恢复至CK水平或超过CK,表现出极强的复水敏感性和潜在恢复能力。该品种扁蓿豆对干旱胁迫及复水的适应主要分为3个时期：主动适应期,其生理参数的可塑性指数为形态参数的1.33倍,主要通过抗氧化及渗透调节来减少水分散失增加水分吸收、缓解氧化伤害以适应干旱逆境;被动适应期,其形态参数的可塑性指数为生理参数的1.31倍,主要采用牺牲生物量的生存策略以及降低色素含量减少光吸收的光保护机制来提高逆境下的生存能力;复水恢复期,根冠比、气孔开放率、气孔及表皮细胞密度比CK分别增加25.9%、29.7%、24.2%和16.3%,其较高的根冠比和叶片较高的气孔开放率及小而密的气孔及表皮细胞特征,保证了直立型扁蓿豆吸水能力以及水分运输效率的迅速恢复。综上,直立型扁蓿豆抗旱能力较强,能够通过形态生理的改变以及调整不同器官的生物量分配来应对与适应干旱逆境及复水,且在不同处理阶段采取不同的适应策略以达到生存目的。  相似文献   

Influence of P and K fertilization on Phytophthora root rot or excess soil water injury of alfalfa cultivars     

《Communications in Soil Science and Plant Analysis》2012,43(2):229-243

Abstract

Phytophthora root rot (PRR) is an important disease of alfalfa (Medicago sativa L.) in wet soils. Excess soil water (ESW) stress is required for the successful infection and development of PRR. The effects of P at 0, 40, and 80 μg.g^‐1 soil and K at 0, 120, and 240 μg.g^‐1 soil (in factorial combinations) on the severity of PRR or ESW stress was investigated on a PRR‐susceptible (Iroquois) and a resistant (Oneida) alfalfa cultivar in a greenhouse trial. P and K applications significantly increased the dry weight of tops prior to imposition of PRR or ESW stress. However, plants subjected to PRR or ESW stress did not respond significantly to P or K and there was no significant effect of P or K on the severity of PRR or ESW injury. The plants subjected to PRR or ESW stress had lower tissue P concentration than those under stress‐free conditions. Increasing P additions resulted in a significant increase in P concentration of Iroquois plants, while in Oneida the response was non‐significant. The response of K concentration in the plant to increasing K additions was dependant on P levels.  相似文献   

新牧一号苜蓿MvP5CS基因的克隆和功能分析   总被引：1，自引：0，他引：1  

张桦  张富春  曾光  张博  陈全家 《草业科学》2012,29(1):51-58

为研究苜蓿（Medicago sativa）耐盐分子机制，为抗逆分子育种提供依据，通过同源克隆和RT PCR技术克隆了新牧一号杂花苜蓿（M.varia Xinmu 1）的MvP5CS基因， MvP5CS全长2 148 bp，Genbank登录号为：EU371644。荧光定量PCR分析发现，在盐胁迫下MvP5CS基因表达明显上调，说明 MvP5CS与苜蓿的耐盐性相关。利用农杆菌介导的叶盘转化法，将MvP5CS基因成功转入烟草（Nicotiana tabacum），盐胁迫下转MvP5CS基因T1代烟草萌发率和根长均高于非转基因烟草。表明新牧一号苜蓿P5CS基因能够提高转基因烟草的耐盐性。  相似文献   

EFFECTS OF pH ON GROWTH AND NODULATION OF TWO FORAGE LEGUMES     

《Journal of plant nutrition》2013,36(9):1367-1375

A study was conducted to compare the responses to acid pH of Medicago sativa and Lotus glaber, two forage legumes with different environmental requirements, either supplied with inorganic nitrogen (N) or inoculated with different strains of their nodule bacteria. Medicago sativa showed, in both treatments, a significant reduction in total dry weight at pH below 6.0. In contrast, Lotus glaber grew equally well at all the pHs assayed in the presence of adequate N. Under inoculated conditions, in the absence of N supply, plant growth was dependent on the bacterial strain used. When the ability of each strain to multiply in culture medium was examined, it was observed that Sinorhizobium meliloti strains showed a pH-sensitive response that inhibited growth at pH 4.0, whereas Mesorhizobium loti strains showed normal growth at this pH. These results suggest that for the effectiveness of Mesorhizobium loti–Lotus glaber symbiosis in acid soils the major factor to be considered is the tolerance of the bacterial strain to acid conditions, while the limiting factors for the Sinorhizobium meliloti–Medicago sativa symbiosis are the sensitivity to low pH of both the plant and its bacteria.  相似文献   

Effectiveness of Coniothyrium minitans and Trichoderma atroviride in suppression of sclerotinia blossom blight of alfalfa   总被引：3，自引：0，他引：3  

G. Q. Li  H. C. Huang †  S. N. Acharya  R. S. Erickson 《Plant pathology》2005,54(2):204-211

The effects of the mycoparasites Coniothyrium minitans and Trichoderma atroviride on the suppression of alfalfa blossom blight caused by Sclerotinia sclerotiorum were evaluated under indoor and field conditions. When T. atroviride (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) were applied to detached young alfalfa florets, T. atroviride effectively inhibited saprophytic growth of S. sclerotiorum, whereas C. minitans showed no inhibition under the same conditions. When T. atroviride (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) was applied to young alfalfa petals in vivo just after pollination, the percentage of pod formation was higher for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum, and the percentage of pod rot was lower for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum. However, when they were applied to senescent petals attached to developing pods of alfalfa at 9·2 × 10⁴ conidia/floret together with S. sclerotiorum at 4·5 × 10³ ascospores/floret at 14 days after pollination, C. minitans was more effective than T. atroviride in suppressing sclerotinia pod rot and seed rot of alfalfa. Field experiments showed that three applications of C. minitans (5·4 × 10⁶ conidia mL⁻¹) or T. atroviride (5·4 × 10⁶ conidia mL⁻¹) at a 7-day interval to blossoms of alfalfa effectively suppressed sclerotinia pod rot in two out of three annual trials. Coniothyrium minitans effectively suppressed sclerotinia seed rot in all three years, whereas T. atroviride was not effective against seed rot in any of the trial years. The efficacy of C. minitans was not significantly different (P > 0·05) from benomyl (250 µg ai mL⁻¹). This study suggests that C. minitans has potential as a biocontrol agent to control blossom blight of alfalfa caused by S. sclerotiorum.  相似文献   

内蒙古中东部野生扁蓿豆种子硬实性的研究   总被引：1，自引：0，他引：1  

额尔敦嘎日迪  中田昇 《中国草地学报》2007,29(6):1-5

扁蓿豆是抗逆性极强的优良牧草,但其种子发芽率低,具有65 %～75 %的硬实。在恒温25℃时发芽率较高,在变温20 ～30℃时发芽率最高,若温度超过30℃或低于15℃时发芽被抑制,但短时间高温处理有利于促进发芽。采用机械或浓硫酸处理,发芽率分别比对照高40.6 %和45.6 %。野生种子的硬实率与采集地气温,特别是与7月份温度有着很高的正相关关系,而且不同产地的种子表现出较大的差异,以赤峰市北部采到的居群硬实率最高,比呼和浩特市采集的居群高9 %。随着储藏年限的推移,种子的硬实率不断下降,发芽率增加,储藏3.5年的种子发芽率达到了67.0 %,表明扁蓿豆种子采集后通过室温储藏可以达到提高发芽率的效果。  相似文献   

环峡南苜蓿种子生产技术研究     

袁福锦薛世明  罗在仁肖院先 《四川草原》2006,(3):10-13

对影响环峡南苜蓿种子产量的磷肥、钾肥、浇水和地膜条件4个因素进行正交组合试验,结果表明: 磷肥、钾肥、磷钾肥互作对种子产量的影响较大,其次为地膜条件、磷肥浇水互作,浇水的影响最小。试验结果和生产成本综合考虑,最适宜的处理组合为:钙镁磷450 kg/hm2+硫酸钾150 kg/hm2+不铺地膜+不浇水。  相似文献