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somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L.‘Moldova’

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Forestry Studies in China 2008年第4期10卷 253-258页

作者： LI Hui LI Feng-lan DU Jing-chuan LU Hai HE Zheng-quan College of Biological Sciences and Biotechnology Beijing Forestry University Beijing 100083 P. R. China Biotechnology Research Center Three Gorges University Yichang 443000 P. R. China

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇（Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.

关键词： zygotic embryo somatic embryo histology Vitis vinifera L.

Thansgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system

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Cell Research 2001年第2期 156-160页

作者： DENG XIANG YANG, ZHI MING WEI, HAI LONG AN Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy Sciences, Shanghai 200032, China 中科院上海植物生理生态所上海 200032

After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

关键词： Peanut (Arachis hypogaea) somatic embryo hygromycin resistance particle bombardment.

Regulation Culture on Cytological, Biochemical and Physiological Characteristics of somatic Carrot embryos

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Developmental and Reproductive Biology 1994年第1期3卷 42-49,T001页

作者：黄美娟黄绍兴朱溦

Cytological,biochemical and physiological charcteristics of quiescent,high-vigor carrot somatic embryos obtained with the use of regulation culture were studied. As shown in cytological observation, quiescent somatic embryos obtained by regulation culturing for 70 days displayed small compacted cells with thick wall, dense cytoplasm and large amount of starch granules. Biochemical tests revealed a starch content and a soluble sugar content 1.9 and 1.7 times respectively as much as those of the control embryos(normal culture) while physiological study demonstrated that the respiration was maintained stally at a very low level,only 1/4 that of the control at the end of the 70-day period. These results provide further evidences that regution culture so after the morphology, biochemistcy, physiologr and metabolism of somatic embryos that they became quiescent,highly synchronized, and had their roots ungrown, a sate facilitating safe storage.

关键词： Carrot somatic embryo Regulation culture Artificial seed.

Precocious In-Vitro Flowering of Perennial Asparagus (Asparagus officinalis L.) Regenerants with a Chemical Inducer

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American Journal of Plant Sciences 2016年第14期7卷 1834-1845页

作者： Fuminori Komai Yasuki Watanabe Akira Kanno Kiyoshi Masuda Graduate School of Agriculture Saga University Saga Japan Graduate School of Life Sciences Tohoku University Sendai Japan Graduate School of Agriculture Hokkaido University Sapporo Japan

A precocious flowering system of regenerants in asparagus (Asparagus officinalis) was achieved by treatment with a chemical inducer. somatic embryos withered completely by being processed for 8 - 12 days with 200 μM n-propyl N-(3,4-dichloro-phenyl)carbamate that had been dissolved in distilled water. In contrast, precocious flowering occurred at an extremely low rate (3.4%) when somatic embryos were processed in carbamate dissolved in Murashige and Skoog’s liquid medium. To encapsulate the female and male embryos, we surveyed the optimum conditions of viscosity and concentration of sodium alginate for encapsulating the seeds, and we screened the values of 80 - 120 cps and 2% - 3%, respectively. The synthetic seeds produced also withered when they were processed with the carbamate dissolved in distilled water. However, when Murashige and Skoog’s liquid medium was used for the solvent, the flowering frequency of the synthetic seeds was enhanced (13.3%). Based on our morphological and histological observations, female and male regenerants that were processed with the carbamate solution produced individual flower organs. The conversion of sex expression did not occur. A precocious flowering system would allow a significant reduction in the time required for perennial seedlings to flower and can, therefore, save time required for further experiments that employ floral homeotic mutants.

关键词： Asparagus Carbamate Encapsulation Precocious Flowering somatic embryo

Plasmolysis treatment enhances the expression of callose synthase gene in zygotic embryos of Eleutherococcus senticosus

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Journal of Forestry Research 2010年第2期21卷 189-192,I0006页

作者：锡林呼安艳夏德安由香玲林木遗传育种与生物技术教育部重点实验室(东北林业大学) 哈尔滨150040

In previous study we reported that pretreatment with plasmolysis enhanced somatic embryo formation in hypocotyls of Eleutherococcus *** the present study,the expression level of callose synthase gene in embryos of *** in response to 2,4-D,sucrose and mannitol treatments was analyzed by *** results show that plasmolysis pretreatment using sucrose and mannitol significantly promoted the expression of callose synthase ***,the thicker cell walls of explant plasmolyzed compared with controls were observed during the somatic *** suggest that the callose may make the cells in epidermis separate from neighboring cells and then develop into embryogenic potential cells.

关键词： Eleutherococcus senticosus somatic embryo Plasmolysis Callose synthase gene RT-PCR