枯斑盘多毛孢菌生物学特性的研究

本文研究了影响松赤枯病病原菌枯斑盘多毛孢菌生长及产孢的因子.枯斑盘多毛孢菌可在较宽的温度范围内生长,自然光照、空气充足有利于产孢;该菌在改良的察氏培养基1号上在25℃恒温条件下菌丝生长速度最快,在改良的察氏培养基2号上产孢量最大且培养8天即可获得最大产孢量,7.47×107个孢子/皿.     

根癌农杆菌介导的淡紫拟青霉遗传转化体系的建立

基于根癌农杆菌介导的遗传转化方法,选择植物线虫生防真菌淡紫拟青霉的分生孢子为受体,建立以G418为筛选标记遗传转化体系.通过优化遗传转化条件,达到较高的转化效率:1000～2400个转化子·10-6分生孢子.对转化子进行PCR验证,表明T-DNA已整合到淡紫拟青霉的基因组中,表型测定发现转化子的遗传表现稳定.农杆菌介导淡紫拟青霉突变体库的建立,为进一步筛选对植物线虫致病性高的突变体、了解淡紫拟青霉的侵染过程和侵染机制提供基础,对培育更优良的植物线虫生防菌株,开发高效生防制剂等具有重要的意义.     

花椒盘二孢生物学特性的研究

花椒盘二抱为花椒褐斑病的病原菌，是作者近年发现的新种。经室内培养试验，明确该病原菌菌落生长及及分生抱子萌发的适宜条件为：菌落在ＰＤＡ培养基上生长较好，温度１０～３０℃均可，２５℃最适，好气，ｐＨ４．５～５．０为宜，碳源以蔗糖为好，氮源以蛋白胖为好。分生孢子发芽温度为１０～３０℃，２０℃最适，需在水滴或相对湿度１００％下才能发芽，ｐＨ４～６为宜，不需营养，光照影响不大。由以上结果可了解该病的有关发病规律，并作为防治研究的依据。     

利用根癌农杆菌转化木本植物的研究进展

本文着重论述了近年来国内外利用根癌农杆菌介导法转化林木的研究进展。简述了获得转基因林木的应用现状,并将国内外林木转基因工作给以概括,提出了转化过程中存在的问题,并对这一转化方法的应用前景进行了展望。     

枯斑盘多毛孢菌粗毒素的基本性质研究

本文对松赤枯病菌（Pestalotia funeren Desm.)培养液粗提物的基本性质进行了初步研究。结果表明，松赤枯病菌培养液中有致病物质即毒素存在，该菌在PD培养液中能达到最大产毒量；松针浸出汁对毒素的产量和致病活性没有明显的促进作用，培养15天的培养液有较高的致萎蔫的活性，随着培养时间的延长其致萎活性没有较大的提高。该毒素是一类极性较大的非蛋白类物质，能用甲醇将其从培养液中较好地提取出来。pH值对毒素粗提液致病活性无显著影响，毒素粗提液中活性成分能被活性碳吸附。     

提高根癌农杆菌介导的香石竹遗传转化效率的研究

以无菌苗叶片为外植体,在根癌农杆菌介导下建立并优化了香石竹5个品种的遗传转化体系。预培养2d可明显提高转化率;香石竹品种间在转化上存在差异;培养基中添加20μmol的AgNO3抑制不定芽的分化。转化植株在含25mg·L-1卡那霉素的生根培养基上培养,生根率为72 1%,GUS检测结果55%的转化植株呈蓝色,PCR扩增表明阳性率为32 2%,Southern杂交证实外源基因已整合到植物基因组中。     

不同环境条件对虫生广布拟盘多毛孢退化的影响

为了解决虫生广布拟盘多毛孢(Entomogenous Pestalotiopsis disseminata)GXSTYJ03菌株在研究和应用中的退化问题,保持菌种稳定性,分别在4种光照(自然光、3天黑暗、24 h黑暗、24 h光照)条件下培养和在3种培养基(PSA、PPSA1、PPSA2)上继代,观察菌落形态和测定菌种生长速率及产孢量,探究不同环境条件对菌种退化的影响。结果表明,环境条件的改变,可导致GXSTYJ03菌株菌落形态局变、生长速率和产孢量的变化。以24 h全光照处理、PPSA1培养基培养可以保持GXSTYJ03菌株的稳定性和减缓其退化能力,并提出了控制菌种退化的措施。     

重寄生拟盘多毛孢cr013菌株聚酮合酶基因的多样性分析

本研究通过基因组挖掘的方式首次从重寄生拟盘多毛孢cr013菌株中获得了聚酮合酶(polyketide syn-thase,PKS)基因,并利用NCBI、CDD、Clustal W、MEGA 7.0等生物信息软件对获得的PKS基因进行功能预测,挑选相似性较高、结构和功能已知的非还原型crPKS3基因和还原型crPKS12...     

杉木球果果鳞受顶枯拟盘多毛孢侵染后细胞结构变化的电镜观察

杉木球果受顶枯拟盘多毛孢（Ｐｅｓｔａｌｏｔｉｏｐｓｉｓａｐｉｃｕｌａｔｕｓ（Ｈｕａｎｇ．）Ｈｕａｎｇ．）侵染后,罹病果鳞细胞的超显微结构依侵染程度不同表现一系列病理变化,主要表现在：叶绿体的基粒片层和膜结构遭到破坏;线粒体内嵴结构和膜结构遭到破坏;基质的许多细胞器解体消失。     

Agrobacterium tumefaciens-mediated GUS gene transfer to Sophora japonica L.

Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefaciens strain LBA4404 that harbored the binary vector pBI121 containing genes forβ-glucuronidase (GUS) and neomycin phos-photransterase (nptⅡ). S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets from the explants. The transformed plants resembled their parents in morphology.     

Agrobacterium tumefaciens-mediated GUS gene transformation of Robinia pseudoacacia ''''Idaho''''

Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia 'Idaho') mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays ofβ-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blotting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia 'Idaho' mediated with A. tumefaciens.     

农杆菌介导高羊茅基因转化体系的建立

以高羊茅(FestucaarundinaceaSchrebb)品系宾哥(Bingo)为材料,建立了农杆菌介导的基因转化方法。高羊茅种子在9mgL-12,4-D作用下,获得胚性愈伤组织。愈伤组织经农杆菌侵染,在30和50mgL-1的潮霉素浓度梯度下筛选,得抗性愈伤组织。组织化学染色法证实,uidA基因在抗性愈伤组织中表达:抗性愈伤组织在分化培养基中分化得到转基因植株。PCR、Southern杂交法证实外源基因已导入到高羊茅植株中。图5参37。     

Induction of tumours by various strains of Agrobacterium tumefaciens on Abies nordmanniana and Picea abies

Abies nordmanniana and Picea abies seedlings were inoculated with a wild‐type (C 58) or an attenuated strain (rooter or shooter mutants) of Agrobacterium tumefaciens. Large tumours were formed on A. nordmanniana in response to the wild‐type and rooter strains, 60–65 % of the seedlings being susceptible. Smaller tumours were formed on Picea abies in response to the wild‐type strain, at least 12% of the seedlings being susceptible. Tumours from both species induced by the wild‐type strain grew in vitro on medium without added phytohormones.     

抗生素对农杆菌的抑制和杨树叶片分化的影响

中嘉8号杨为湖北省林业厅和中国林科院从美洲黑杨的实生后代中选育的杨树新品系。以中嘉8号杨为材料,探讨了几种抗生素对农杆菌的抑制和对杨树离体叶片分化的影响。结果表明,噻孢霉素的抑菌效果最好,选用100~200 mg/L的噻孢霉素来抑菌是比较合适的,羧苄青霉素和链霉素对杨树叶片离体培养的毒性比噻孢霉素大,卡那霉素完全抑制芽的分化。     

根癌农杆菌介导Caz基因转化烟草条件研究

利用根癌农杆菌介导Caz基因转化烟草,将外源基因导入烟草愈伤组织,建成可诱导表达细胞质和细胞核CaM/CaM结合多肽的转基因烟草．研究结果表明:植物材料的选择和培养、农杆菌的培养和稀释、植物材料与农杆菌共培养的程度及转基因植株的筛选条件是影响农杆菌转化效率的重要因素．     

Regeneration and gene transformation systems of Robinia pseudoacacia ''Idaho'' mediated by Agrobacterium tumefaciens

Robinia pseudoacacia 'Idaho' is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L-1.     

Agrobacterium tumefaciens-mediated GUS gene transformation of Robinia pseudoacacia ‘Idaho’

Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia ‘Idaho’) mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays of β-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blot-ting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia ‘Idaho’ mediated with A. tumefaciens.