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Pedosphere 2005年第3期15卷 281-285页

作者： ZHAOYan-Wen HUZheng-Yi CAOZhi-Hong J.D.BEATON A.M.HENDERSON M.X.FAN XUCheng-Kai InstituteofSoilScience ChineseAcademyofSciencesNanjing210008（China） FernzSulFerWorksInc. P.O.Box9IrricanaAB.TOM1BO（Canada） TheSulphurInstitute WashingtonDC20036（USA） CollegeofAgriculture NanjingAgri.Univ.KeyLab.ofCropGrowthRegulationMinistryofAgri.Nanjing210095（China）／／InstituteofSoilScienceChineseAcademyofSciencesNanjing210008（China）

Cocoon samples were collected from fifty-two mulberry gardens with high, intermediate, and low silkworm cocoon productivities in the lower-middle reaches of the Yangtze River in the six China’s provinces of Jiangsu, Jiangxi, Anhui, Fujian, Hunan, and Hubei to determine the transformation efficiency of S from mulberry leaves to silkworm cocoons, and to evaluate the sulfur cycle (uptake and output) in the mulberry leaf-silkworm cocoon system with typical mulberry gardens in the lower-middle reaches of the Yangtze River in China. The transformation efficiency of sulfur (TES) from mulberry leaves into silkworm cocoons in the high-productivity mulberry gardens was significantly lower (P < 0.05) than that in the low-productivity gardens. For the high-productivity mulberry gardens the TES from mulberry leaves into the cocoon shells was significantly higher (P < 0.05) than that for low-yield mulberry gardens. Producing 1 kg dry cocoon in mulberry gardens required uptake of about 20 g S, however 1 kg of dry cocoon only removed about 4 g S. Therefore, recycling of these organic wastes with silkworm cultivation was important for sulfur balances.

关键词： mulberry leaves silkworm cocoon sulfur transformation efficiency

Comparison of transformation efficiency of piggyBac Transposon among Three Different Silkworm Bombyx mori Strains

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Acta Biochimica et Biophysica Sinica 2007年第2期39卷 117-122页

作者： Boxiong ZHONG~1 Jianying LI~1 Jin'e CHEN~(1,2) Jian YE~1 and Songdong YU~1 ~1College of Animal Sciences,Zhejiang University,Hangzhou 310029,China ~2Institute of Sericultural Research,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China College of Animal Sciences Zhejiang University Hangzhou 310029 China Institute of Sericultural Research Zhejiang Academy of Agricultural Sciences Hangzhou 310021 China

The transformation rate of three different strains of silkworm Bombyx mori was comparedafter the introduction of enhanced green fluorescence protein （EGFP）-encoding genes into the silkwormeggs by microinjection of a mixture of piggyBac vector and helper plasmid containing a *** there were no significant differences among the three strains in the percentages offertile moths in microinjected eggs （P=0.1258）,the percentages of G0transformed moths in fertile mothsand injected eggs were both significantly different （P=0.01368 and P=0.02398, respectively）.Thetransformation rate of the Nistari strain （Indian strain） was significantly higher than that of the other twostrains,Golden-yellow-cocoon （Vietnamese strain） and Jiaqiu （Chinese strain）,which had similar rate. Theseresults indicate that the transformation efficiency of the piggyBac-based system might vary with silkwormstrains with different genetic *** presence of endogenous piggyBac-like elements might bean important factor influencing the transformation efficiency of introduced piggyBac-derived vectors,andthe diverse amount and activation in different silkworm strains might account for the significant differences.

关键词： Bombyx mori piggyBac transposon transgenesis transformation efficiency

Effect of Vector Density and Competence on Macromolecular Vector transformation efficiency

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Asian Agricultural Research 2016年第12期8卷 81-84,88页

作者： Hongni Qin Lanlan ZHANG Suzhou Industrial Park Institute of Services Outsourcing Genewiz (Suzhou) Biotechnology Co. Ltd.

To research the effects of vector quantity and competence on the positive cloning rate,with a known gene sequence but in the absence of DNA template,we artificially designed 26 primers to synthesize a target gene of 835 bp in vitro using overlapping PCR technique. The whole experiment design with two factors and six levels( 36 combinations) was applied to study the effects of the vector density and competent cells on the macromolecular vector transformation efficiency. Based on the 1 500 ng target gene,the vector density grades were designed( 50,100,150,200,250,300 ng),and then the recombinant plasmids were transformed into Top10F',DH5,Stbl3,Epi400,JM108,SCSI. Results showed that the positive cloning rates of different vector amount from big to small were in the order of 200,250,300,150,100 and 50 *** maximum positive cloning rate of 200 ng reached 75%; and the average value was 28. 5%. The positive cloning rates of different competent cells from big to small were in the order of stbl3,Top10F',DH5,JM108,Epi400 and SCSI. Stbl3 was higher than other competent cells under any vector density,and its average positive cloning rate was 42. 4%. Both the vector density and competent cells had significant effects on the macromolecular vector transformation efficiency. The optimal combination was C4 with 200 ng vector density and Stbl3,the positive cloning rate of which could reach 75%.

关键词： Macromolecular vector Vector density Competence transformation efficiency

Effects of the Ice Bath Time after Heat Shock and the Incubation Time on the efficiency of Chemical transformation Method

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Agricultural Science & Technology 2010年第9期11卷 198-200页

作者：郭姣洁薛永常薛张伟徐书景鞠建松大连工业大学生物工程学院辽宁大连116034 河北师范大学生命科学学院河北石家庄050016

[Objective] The aim of the research was to study the effect of the ice bath time after heat shock and the incubation time on the transformation efficacy,and to establish a simple and quick transformation method.[Method]Competent cells were prepared with two buffer solutions;with the ice bath time after heat shock and the recovery time as the variables,the relationship between these two factors and transformation efficacy was studied.[Result]The transformation efficacy was the best when the ice bath time was 2 min and the recovery time was 30 or 40 min;when the ice bath time and the recovery time was 0 min,a certain amount of transformants still could be obtained.[Conclusion]The ice bath time after heat shock and the recovery time had certain impact on transformation efficacy,but they were not the decisive ***,in the general transformation experiment,these two steps could be omitted.

关键词： Competent cell transformation efficiency Ice bath time Recovery time

Fast,simple,efficient Agrobacterium rhizogenes-mediated transformation system to non-heading Chinese cabbage with transgenic roots

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Horticultural Plant Journal 2024年第2期10卷 450-460页

作者： Huiyu Wang Yushan Zheng Qian Zhou Ying Li Tongkun Liu Xilin Hou State Key Laboratory of Crop Genetics&Germplasm Enhancement Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(East China)Ministry of Agriculture and Rural Affairs of ChinaEngineering Research Center of Germplasm Enhancement and Utilization of Horticultural CropsMinistry of Education of ChinaNanjing Agricultural UniversityNanjingJiangsu 210095China Nanjing Suman Plasma Engineering Research Institute Nanjing Agricultural UniversityNanjingJiangsu 210095China

Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage. Agrobacterium rhizogenes-mediated(ARM) transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants. Here, we optimized conventional ARM approaches(one-step and two-step transformation methods) suitable for living non-heading Chinese cabbage plants in nonsterile environments. Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection, fluorescence observation, and PCR analysis. The transformation efficiency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing, Huangmeigui, Wuyueman, and Sijiu Caixin) was 43.33%-51.09%, whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88% for the 30-day-old Sijiu *** one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60%, and both methods can be used to obtain transgenic roots for functional studies within one month. Finally, optimized ARM transformation methods can easily,quickly, and effectively produce composite non-heading Chinese cabbage plants with transgenic roots, providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.

关键词： Agrobacterium rhizogenes Non-heading Chinese cabbage Transgenic roots Composite plant transformation efficiency

Optimization of protoplast isolation,transformation and its application in sugarcane (Saccharum spontaneum L.)

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The Crop Journal 2021年第1期9卷 133-142页

作者： Qiongli Wang Guangrun Yu Zhiyong Chen Jinlei Han Yufang Hu Kai Wang Key Laboratory of Genetics Breeding and Multiple Utilization of CropsMinistry of Education/Fujian Provincial Key Laboratory of Haixia Applied Plant Systems BiologyCenter for Genomics and BiotechnologyFujian Agriculture and Forestry UniversityFuzhou 350002FujianChina National Engineering Research Center of Sugarcane Fujian Agriculture and Forestry UniversityFuzhou 350002FujianChina

Sugarcane is a prominent source of sugar and ethanol *** analysis for trait improvement of sugarcane is greatly hindered by its complex genome,long breeding cycle,and recalcitrance to genetic *** protoplast-based transient transformation system is a versatile and convenient tool for in vivo functional gene analysis;however,quick and effective transformation systems are still lacking for ***,we developed an efficient protoplast-based transformation system by optimizing conditions of protoplasts isolation and PEG-mediated transformation in *** yield of viable protoplasts was approximately 1.26×107 per gram of leaf material,and the transformation efficiency of 80.19%could be achieved under the optimized ***,using this approach,the nuclear localization of an ABI5-like bZIPs transcription factor was validated,and the promoter activity of several putative DNase I hypersensitive sites(DHSs)was *** results indicated this system can be conveniently applied to protein subcellular localization and promoter activity assays.A highly efficient *** mesophyll cell protoplast isolation and transient transformation method was developed,and it shall be suitable for in vivo functional gene analysis in sugarcane.

关键词： Saccharum spontaneum L. Protoplast isolation PEG transformation efficiency Transient expression

Efficient expression of green fluorescent protein (GFP) mediated by a chimeric promoter in Chlamydomonas reinhardtii

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Chinese Journal of Oceanology and Limnology 2008年第3期26卷 242-247页

作者：吴锦霞胡章立王潮岗黎双飞雷安平 Institute of Ecology and Environment College of Life Sciences Shenzhen UniversityShenzhen 518060 China

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.

关键词： expression efficiency green fluorescent protein (GFP) HSP70A-RBCS2 RBCS2 transformation efficiency

Establishment of a transformation system in close relatives of wheat under the assistance of TaWOX5

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Journal of Integrative Agriculture 2024年第6期23卷 1839-1849页

作者： Yanan Chang Junxian Liu Chang Liu Huiyun Liu Huali Tang Yuliang Qiu Zhishan Lin Ke Wang Yueming Yan Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China Beijing Key Laboratory of Plant Gene Resources and Biotechnology for Carbon Reduction and Environment Improvement/College of Life Science Capital Normal UniversityBeijing 100048China College of Agronomy and Biotechnology China Agricultural UniversityBeijing 100193China State Key Laboratory of Plant Cell and Chromosome Engineering/Institute of Genetics and Developmental Biology Chinese Academy of SciencesBeijing 100101China

Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat *** this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature *** plants were efficiently *** the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar *** transformation efficiency of *** genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic *** and hexaploid triticale *** the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR ***,the transgene was almost stably inherited in the next generation by Mendel’s *** findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.

关键词： Triticum monococcum hexaploid triticale rye TaWOX5 Agrobacterium transformation efficiency

Development of powdery mildew resistant derivatives of wheat variety Fielder for use in genetic transformation

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The Crop Journal 2023年第2期11卷 573-583页

作者： Xiaona Liang Xiaomin Bie Yuliang Qiu Ke Wang Zujun Yang Yuqi Jia Zhiying Xu Mei Yu Lipu Du Zhishan Lin Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China College of Life Science Shandong Agricultural UniversityTai’an 271018ShandongChina School of Life Science and Technology University of Electronic Science and Technology of ChinaChengdu 611731SichuanChina College of Agriculture Yangtze UniversityJingzhou 434025HubeiChina National Key Facility of Crop Gene Resources and Genetic Improvement Beijing 100081China Key Laboratory of Ministry of Agriculture and Rural Affairs of China for Biology and Genetic Breeding of Triticeae Crops Beijing 100081China

Genetic transformation is widely used to improve target traits and to study gene function in ***,transformation efficiency depends on the physiological status of the recipient genotype and that is affected by several factors including powdery mildew(PM)*** widely used recipient variety Fielder is very susceptible to ***,it would be beneficial to develop PM resistant derivatives with high regeneration ability for use in genetic *** the present study PM resistant lines CB037 and Pm97033 carrying genes Pm21 and PmV,respectively,were backcrossed to Fielder with selection for PM *** lines,NT89,NT90,NT154,and WT48 with Pm21 and line FL347 with PmV were developed,identified by molecular markers and genomic in situ hybridization(GISH)or fluorescent in situ hybridization(FISH),and further subjected to detailed assessment of agronomic traits and regeneration ability following genetic transformation *** FL347,WT48,NT89 and NT154 assessed as being equal to,or superior,to Fielder in regeneration and transformation ability are recommended as suitable materials for the replacement of Fielder for wheat gene transfer and genome editing study.

关键词： Genotype improvement Plant regeneration ability transformation efficiency Triticum aestivum

Optimization of Electroporation Processing of Bacillus subtilis

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Optimization of Electroporation Processing of Bacillus subti...