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Phytosulfokine(PSK)is a new peptide plant hormone,which was isolated in the conditioned medium of cultures derived from both monoco-tyledonous and dicotyledonous plants,such as Asparagus officinalis mesophyll[1],rice[2],zinnia[3],and carrot[4].PSK has two types of structure:PSK-αand PSK-β.The former is a sulfated pentapeptide[H-Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln-OH],the latter is a sulfated terapeptide[H-Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-OH].Both are heat-stable,susceptible to pronase… 相似文献
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以香蕉(Musa spp.)品种‘天宝蕉’(Musa spp.,AAA类群)为试材,对以根癌农杆菌介导法的香蕉遗传转化体系进行较全面的研究,并以该系统进行了ACS反义基因转化香蕉的研究。结果表明:不经预培养的香蕉茎尖横切薄片,侵染前用附加0.1 mg/L甘露醇的高渗固体培养基前处理4 h,农杆菌重悬液浓度为OD600在1.0左右,重悬液中含100 g/L蔗糖,接菌时间为10~15 min,于26℃黑暗条件下共培养4 d,共培养培养基pH值为5.8是较为适合的转化条件;采用附加100 mg/L卡那霉素、2 mg/L AgNO3筛选培养基对共培养后的香蕉横切薄片进行筛选,共获得5个转ACS反义基因的抗性芽系;经GUS组织化学法及PCR检测,gus基因已整合进香蕉基因组。 相似文献
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Thi Thanh Nhan Pham Thi Ngoc Lan Nguyen Thi Ha Bui Huu Quan Nguyen Thi Tam Nguyen Van Son Le 《The Journal of Horticultural Science and Biotechnology》2019,94(5):591-598
Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg?1 to 5.12 μg.mg?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g?1 (fresh weight) to 10.53 µg.g?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter 相似文献
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甘薯SAAT法遗传转化条件的优化 总被引:4,自引:0,他引:4
SAAT即声波辅助的农杆菌遗传转化方法。在遗传转化中,超声波处理可以在外植体上产生许多易于农杆菌进入的微小伤口和通道,从而提高转化率。该研究首次将SAAT法应用于甘薯遗传转化并对遗传转化体系进行优化,根癌农杆菌菌株为EHA101/pROA93(nptⅡ和gus),受体材料为甘薯品种栗子香的胚性悬浮细胞。结果表明,悬浮培养105 d,继代培养3 d,将经超声波处理30 s的胚性悬浮细胞侵染到OD600值稀释至0.5的对数生长期的EHA101菌液中10 m in,经4 d共培养后,GUS瞬时表达率最高,达到86.7%,稳定表达率为37.9%;转化时加入乙酰丁香酮亦能提高转化率;转基因植株筛选的适宜选择压为Kan 10 mg/L;适宜的抑菌剂为Carb 100 mg/L。 相似文献
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黄瓜花叶病毒外壳蛋白基因转化辣椒的研究 总被引:2,自引:0,他引:2
以辣椒品种'B162'为试材,探讨了通过根癌农杆菌介导法将黄瓜花叶病毒外壳蛋白(CMV-CP)基因转化辣椒的适宜条件.结果表明,将辣椒带柄子叶进行预培养2 d后,用D600 nm为0.3的根癌农杆菌菌液浸泡7 min,再经过共培养2 d后转移到筛选分化培养基中进行培养,有利于获得辣椒抗性不定芽.通过对获得的10株抗性不定芽进行PCR检测,其中有2株扩增出目的条带,初步证明CMV-CP基因已经转入辣椒不定芽中. 相似文献
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[目的]将磷酸烯醇式丙酮酸羧化酶(PEPCase)基因导入普通小麦临优145。[方法]利用根癌农杆菌遗传转化系统,以普通小麦临优145为材料,将磷酸烯醇式丙酮酸羧化酶(PEPCase)基因,导入小麦胚性愈伤组织中,用含潮霉素(hyg)的筛选培养基连续筛选,并从分子水平上检测该基因。[结果]获得了hyg抗性植株,经GUS组织化学染色检测表明,抗性苗叶片被染成了深蓝色;PCR检测出目标条带。[结论]初步证明磷酸烯醇式丙酮酸羧化酶(PEPCase)基因已经导入受体材料中。 相似文献
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选用高感大豆花叶病毒病而再生能力较强的大豆品种和品系作为受体,对丛生芽诱导培养基、侵染菌液的制备以及根的诱导方法进行了优化。采用优化后的遗传转化体系以农杆菌介导法将pac1导入大豆,MSB培养基中附加2 mg/L6-BA+0.2 mg/LIBA进行丛生芽诱导,液体YEB重悬活化菌体后侵染,MSB培养基中蔗糖含量15%+2 mg/LIBA+0.2 mg/L6-BA进行生根诱导。共获得8株PCR检测呈阳性植株,经SouthernBlot检测,证明pac1基因已整合到大豆基因组中。转基因植株对病毒的抗性评价正在进行中。 相似文献
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影响农杆菌介导甜瓜子叶遗传转化的因素 总被引:4,自引:0,他引:4
以甜瓜子叶作为根癌农杆菌介导转化的受体,通过GUS基因瞬时表达率的分析,研究此转化体系的最佳实验参数.实验结果表明,预培养时间、预培养后对外植体进行处理、感染时间、共培养时间、农杆菌工程菌的浓度等对转化效率都有一定的影响,但是根癌农杆菌诱导物AS并不能大幅度提高转化效果.对外植体进行重新处理,预培养2 d,用OD560为0.3的农杆菌工程菌感染15~25 min,共培养3~4 d的理想条件下,GUS瞬时表达率可达85%. 相似文献