枸杞胚乳植株的研究及应用潜力

本文对10株不同倍性的(2x,3x,4x,混倍体)构杞胚乳植株进行了研究。观察到3x,4x和混倍体植株的气孔、叶片、花药较二倍体枸杞增大。花器有明显的变异现象。育性显著降低,种子中多数是秕粒。3x植株中有二株,平均每个果内仅有2.7和3.3粒饱满种子,比对照分别减少91.8％和89.9％。在3x植株的减数分裂过程中,观察到许多染色体行为异常现象。     

STUDY OF ENDOSPERM PLANTS OF Lycium barbarum L. AND ITS POTENTIALITY FOR PRACTICAL USE

This paper reports morphological differences between diploid, triploid, tetraploid, mixploid endosperm plants and the original diploid plants. In comparison with original diploids, the triploid, tetraploid and mixploid endosperm plants have larger stomas, greater variability in flowor organs, decreased fertility, decreased seed set per fruit, and a lot of abnormalities in chromosome behaviour in meiosis. The propagation of endosperm triploid seedless plants is in progress.     

致瘤农杆菌能够转化大麦和小麦

直到现在,单子叶植物特别是禾谷类作物的基因工程还没有合适的载体系统。本文首次报道了致瘤农杆菌T37,A208和B63个菌种能够感染大麦和小麦的部分品种,形成突起和肿瘤。并且发现,酚类化合物乙酰丁香酮对转化有促进作用。另外,在合适的植物组织接种农杆菌也是转化成功的关键。文中讨论了农杆菌的宿主范围以及Ti质粒作为禾谷类作物基因工程载体的可能性。     

PLANT REGENERATION FROM IMMATURE COTYLEDONOUS TISSUE CULTURES OF SOYBEAN BY SOMATIC EMBRYOGENESIS

Genetic engineering of soybean is limited because of the difficulty of regeneration of plants via in vitro culture. Here we report that a method of obtaining somatic embryos and regenerated plants of soybean has been developed. It is achieved by culturing immature cotyledons of soybean on modified Murashige-Skoog medium supplemented with high concentration of either napthalene acetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). High induction frequencies of 85% and 94% are reached by using 10mg/l NAA and 5mg/l 2,4-D, respectively. A number of regenerated plantlets of soybean are transplanted to sterilized soil and grown into 15 full plants in pots.     

根瘤农杆菌介导的枸杞转化及转化植物的获得

本文报道一种重要的中药——枸杞(Lycium barbarum L.)的转化、再生系统的建立。以根瘤农杆菌(Agrobaterium tumefaciens)C58cl(pGV3850::neo1103)感染枸杞的幼茎外植体,在含卡那霉素50μg/ml的诱培养基上选择转化的愈伤组织,在含卡那霉素25μg/ml分化培养基上得到再生小芽,再生小芽中有30％外观形态正常。形态正常的小芽转到生根培养基后可长出根从而得到完整的植株。胭脂碱检测。NPT-Ⅱ活性测定及分子杂交的结果均表明,外源基因已导入枸杞并得以表达。在实验过程中还发现:(1) 经过一定预处理后,形成愈伤组织较快的外植体易被转化。(2) 转化后愈伤组织再生正常植株的频率比未转化的愈伤组织高。     

Genetic Transformation of Lycium barbarum L. via A. tumefaciens

A system for transformation and regeneration of Lycium barbarum L. , an important Chinese medical plant, has been established. Young stem segments from Lycium barbarum L. were infected with Agrobacterium tumefaciens C58cl(pGV3850: :ineo1130), and the transformed calli selected from the callus induction medium containing 50 fig/ml kanamycin could regenerate buds on differentiation medium containing 25 μg/ml kanamycin. 30% of the regenerated buds were normal in morphology. The normal buds could develop into whole plantlets after they were transferred to the rooting medium to induce roots. Nopa-line detection, NPT-Ⅱ enzyme activity assay and Southern blotting hybridization indicated that the foreign genes had been integrated into the genome of Lycium barbarum L. and expressed in the plant. In the processes of experiments, it was found that (i) after the pre-processes, the explants which formed callus quickly were easy to transform ; (Ⅱ) the rate of normal regenerated plants from transgenic calli was hig     

A STUDY ON TUMOR FORMATION OF SOYBEAN AND GENE TRANSFER

From 15 strains of Agrobacterium tumefaciens 7 were selected as the best inductors oftumors on soybean and 94 tumor formation genotypes were screened from 1553 varieties andforms of Glycine soja, G. gracilis and G. max. The frequency of tumor formation was 6%.The bacteria-free calluses were obtained. The data of biochemical analysis verify that someof the above-mentioned calluses contained nopaline. This means that Ti-plasmid of Agrobac-terium tumefaciens can be used as a vector for the transfer of nopaline, a marker gene, intogenome of several species of soybean. In the soybean genome the transferred nopaline genewas integrated and expressed. The transformed gene can be stably integrated in the soybeangenome through vegetative propagation.     

EXPRESSION OF A ZEIN GENE (Z4) INTRODUCED INTO DICOTYLEDONOUS Solanum nigrum

A maize genomic clone containing a zein gene (Z4) is inserted into the T-DNA of the Ti plasmid pTiT37. Agrobacterium tumefaciens strain harboring this modified Ti plasmid is used to infect stem sections of young plants or explants of dicotyledonous Solanum nigrum. Axenic transformed calli active in nopaline synthesis are obtained and transgenic plants are differentiated from them DNA Southern hybridization and RNA dot-hybridization analyses show that the zein gene is really transferred and integrated into the nuclear genome of transformed Solanum nigrum and that the zein gene can be transcribed into mRNA in the transformed calli and shoots. But the presence of the zein protein cannot be detected in either the transformed calli or the transgenic shoots. The results of thte experiments demonstrate that the promoter of a gene from monocotyledonous plants can function normally in transgenic dicots. The possibility of developmentally-regulated expression of the zein gene in transformed dicots is discussed in     

人参花粉植株的诱导及体细胞无性系的建立

人参花药培养已连续四年(1981—1984年)得到了花粉植株,并建立了体细胞无性系。人参花药对培养基的适应较广,在MS等6种基本培养基上都得到了愈伤组织;低温预处理能明显提高诱导效果;已筛选出愈伤组织诱导率高的培养基配方,再生植株的分化比较困难,在100多种分化培养基配方中筛选出16种能分化再生植株的配方,分化率高的可达6.8％。经二年多16次继代培养,建立了能保持良好分化能力的体细胞无性系。     

TRANSFORMATION OF Lotus corniculatus L.MEDIATED BY Agrobacterium tumefaciens

A simple and effective system for the transformation and regeneration of a leguminous plant has been developed. The cotyledon explants of Lotus corniculatus (var. Leo) were infected by Agrobacterium tumefaciens that contained a non-oncogenic Ti plasmid vector. The vector carried a chimaeric npt-Ⅱ gene and a nopaline synthase gene (nos). On the selective medium that contained kanamycin, 40% of the explants formed buds within 3 weeks. Developed shoots were cut off and transferred to rooting medium. Normal-looking plants were recovered and grew well after being transplanted into soil, bloomed and set seeds. Foreign genes were integrated onto the L. corniculatus genome, expressed, and inherited through sexual reproduction, which was proven by nopaline detection, NPT-Ⅱ enzyme activity detection and DNA hybridization test of the transformed plants and the nopaline detection of the progeny.