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Medium effects on micropropagation and genetic stability of Citrullus lanatus oleaginous type

Agricultural Sciences 2013年第7期4卷 32-44页

作者： Yah Gwladys Gnamien Irié Arsène Zoro Bi Yatty Justin Kouadio Yves Brostaux Jean-Pierre Baudoin Laboratory of Biology and Improving Vegetable Productions Natural Sciences UnitUniversity Nangui Abrogoua(UNA)AbidjanCote d’Ivoire Tropical Crop Husbandry and Horticulture Gembloux Agro-Bio Tech(GxABT)University of LiègeGemblouxBelgium Department of Applied Statistics Computer Science and MathematicsGembloux Agro-Bio Tech(GxABT)University of LiègeGemblouxBelgium

To regenerate adventitious shoots from the cotyledon proximal parts of Citrullus lanatus (Thunb.) Matsum. and Nakai ssp. mucosospermus (Fursa) oleaginous type, different concentrations of MS mineral elements, sucrose, 6-benzylaminopurine (BAP) and agar were tested. Shoot induction proved to depend on the interaction between levels of sucrose, BAP and MS mineral elements in the medium. The medium containing 3/2 strength of MS mineral elements, 35 g/l sucrose and 1 mg/l BAP solidified with 6 g/l agar allowed the production of numerous shoots without a callus phase. After 3 weeks of culture, 76.7% of the cotyledon proximal parts induced shoots with an average of 12.26 shoots per explant and a mean shoot length of 17.13 mm. The induced shoots were directly rooted and thus complete plants ready for acclimatization were obtained using a two steps procedure. Depending on the genotype, the shoot induction from cotyledon proximal parts ranged from 54% to 96%. Rooted plantlets were acclimatized and transferred to field, where they grew well, developed flowers and fruits like seeded plants. The assessment of the genetic stability of the in-vitro-regenerated plantlets by means of an Amplified Fragment Length Polymorphism (AFLP) analysis with the combination of 5 primers revealed no differences between regenerated plantlets and mother plants.

关键词： AFLP genetic stability In Vitro Oleaginous Seeds Organogenesis

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AFLP Analysis of genetic stability of Sugarcane Tissue Culture Clones

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Agricultural Biotechnology 2020年第1期9卷 1-5页

作者： Fangye LAO Huiyi HE Rui LIU Yongwen QI Guangdong Bioengineering Institute(Guangzhou Sugarcane Industry Research Institute)/Guangdong Key Lab of Sugarcane Improvement & Biorefinery

[Objectives] This study was conducted to better understand the variation law of sugarcane clones during tissue culture process,and to provide a reference for rapid propagation and detection of healthy sugarcane seedlings.[Methods] The genetic stability of tissue culture clones of three sugarcane varieties was analyzed using the AFLP molecular marker technique.[Results]The average number of polymorphic loci was 19. 58 for each primer pair,and the percentage of polymorphic loci was 41. 74%. Compared with the donor varieties,all tissue culture materials were mutated. There were 3-16 missing bands,with an average of 5. 2 bands,and there were 0-17 increased bands,with an average of 3. 3 bands. The total number of missing and increased bands was 4-33,with an average of 8. 5. The band difference rates were in the range of 0. 009 4%-0. 077 6%,with an average of 0. 020 6%. The genetic similarity coefficients between materials ranged from 0. 685 6 to 0. 998 2,with an average of 0. 818 4. The three sugarcane varieties and their tissue culture clones were clustered into three groups.[Conclusions] Although variations occur in tissue culture,the variations are not too obvious,and the genetic stability is relatively high. It is recommended to minimize the number of subculture generations and cultivation time to reduce the occurrence of variation during tissue culture for rapid propagation of sugarcane seedlings.

关键词： Sugarcane Tissue culture Clone AFLP genetic stability

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genetic and agronomic traits stability of marker-free transgenic wheat plants generated from Agrobacterium-mediated co-transformation in T2 and T3 generations

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Journal of Integrative Agriculture 2020年第1期19卷 23-32页

作者： LIU Hui-yun WANG Ke WANG Jing DU Li-pu PEI Xin-wu YE Xing-guo Biotechnology Research Institute Chinese Academy of Agricultural SciencesBeijing 100081P.R.China Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081P.R.China

genetically modified wheat has not been commercially utilized in agriculture largely due to regulatory hurdles associated with traditional transformation methods. Development of marker-free transgenic wheat plants will help to facilitate biosafety evaluation and the eventual environmental release of transgenic wheat varieties. In this study, the marker-free transgenic wheat plants previously obtained by Agrobacterium-mediated co-transformation of double T-DNAs vector were identified by fluorescence in situ hybridization(FISH) in the T1 generation, and their genetic stability and agronomic traits were analyzed in T2 and T3 generations. FISH analysis indicated that the transgene often integrated into a position at the distal region of wheat chromosomes. Furthermore, we show that the GUS transgene was stably inherited in the marker-free transgenic plants in T1 to T3 generations. No significant differences in agronomic traits or grain characteristics were observed in T3 generation, with the exception of a small variation in spike length and grains per spike in a few lines. The selection marker of bar gene was not found in the transgenic plants through T1 to T3 generations. The results from this investigation lay a solid foundation for the potential application of the marker-free transgenic wheat plants achieved through the co-transformation of double T-DNAs vector by Agrobacterium in agriculture after biosafty evaluation.

关键词： wheat marker-free transgenic plants fluorescenee in situ hybridization genetic stability

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Vital Parameters Assessments of Starvation Tolerance of in vitro Populus alba Culture

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Phyton-International Journal of Experimental Botany 2022年第4期91卷 889-902页

作者： Mohamed Fathy Ahmed Eman Zakaria Ahmed Salman Aloufi Mohammed Alqurashi Amal Alyamani Eman Fayad Eman Tawfik Hussien Horticulture Department Faculty of AgricultureAin Shams UniversityCairo11566Egypt Botany and Microbiology Department Faculty of ScienceHelwan UniversityCairo11795Egypt Department of Biotechnology Faculty of SciencesTaif UniversityTaif21944Saudi Arabia

Populus alba is a large woody deciduous *** plant has been introduced to shooting,then multiplication of rooting on Murashige and Skoog(MS)*** work was designed to estimate the effect of two factors(low levels of 1-Naphthaleneacetic acid NAA and sucrose)on *** response resulting in 6 treatments compared to the control,with twelve measured *** was a significant difference in some measurements in morphology,like plantlets fresh-weight,shoot-,root-length,and leaf *** the physiological measurements,there were significant differences in all the measured *** low concentrations of sucrose and media composition/power(MS grams/L)led to starvation in plants;however,these conditions led to enhancement in some morphological and physiological parameters to overcome the starvation effect,compared to the *** RAPD-PCR molecular marker(four decamers)was used to evaluate the new individuals’genetic variation(instability),resulting in a total polymorphism percentage of 50.83%.It was formerly known that the plantlets were identical to each other and to the mother *** this study,however,the use of distinct media power,hormonal and sucrose levels resulted in molecular variation reflected in ***’s morphological and physiological responses.

关键词： genetic stability Populus alba RAPD-PCR somaclonal variations starvation

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Development and Assessment of Two Highly Pathogenic Avian Influenza(HPAI) H5N6 Candidate Vaccine Viruses for Pandemic Preparedness

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Biomedical and Environmental Sciences 2020年第9期33卷 670-679页

作者： LIU Li Qi LI Zi JIAO Ming LU Jian ZHOU Jian Fang LI Xi Yan LIU Jia GUO Jun Feng XIAO Ning ZHAO Xiang WANG Da Yan Chinese National Influenza Center National Institute for Viral Disease Control and PreventionChina CDCBeijing 102206China

Objective In China, 24 cases of human infection with highly pathogenic avian influenza(HPAI) H5 N6 virus have been confirmed since the first confirmed case in 2014. Therefore, we developed and assessed two H5 N6 candidate vaccine viruses(CVVs).Methods In accordance with the World Health Organization(WHO) recommendations, we constructed two reassortant viruses using reverse genetics(RG) technology to match the two different epidemic H5 N6 viruses. We performed complete genome sequencing to determine the genetic stability. We assessed the growth ability of the studied viruses in MDCK cells and conducted a hemagglutination inhibition assay to analyze their antigenicity. Pathogenicity attenuation was also evaluated in vitro and in *** The results showed that no mutations occurred in hemagglutinin or neuraminidase, and both CVVs retained their original antigenicity. The replication capacity of the two CVVs reached a level similar to that of A/Puerto Rico/8/34 in MDCK cells. The two CVVs showed low pathogenicity in vitro and in vivo, which are in line with the WHO requirements for *** We obtained two genetically stable CVVs of HPAI H5N6 with high growth characteristics,which may aid in our preparedness for a potential H5N6 pandemic.

关键词： Highly pathogenic avian influenza H5N6 virus genetic stability Candidate vaccine virus Reverse genetic technology

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Identification and characterization of the new-type fowlpox virus shuttle vectors

Identification and characterization of the new-type fowlpox ...

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第九届全国免疫学学术大会

作者： Shouwen Du Cunxia Liu Yilong Zhu Dayong Ren Maopeng Wang Peng Tan Xiao Li Mingyao Tian Chang Li Ningyi Jin Key Laboratory of Jilin Province for Zoonosis Prevention and Control Institute of Military Veterinary AMMS

Background and Objective:Viral vector is an important vehicle in the field of vaccine ***(APVs)including fowlpox virus(FPV)exhibit strict host range restriction to avian species,and undergo abortive infection in mammalian cells,but correctly express the foreign genes in mammalian cells,and elicit a long-lasting immune response in the vaccinated *** characteristics and their ability to accommodate multiple gene inserts make APVs promising gene therapy and vaccine vectors for diseases caused by pathogenic microorganisms and ***,we parsed the whole-genome information of FPV 282E4 strain,and based on it,the suitable insertion sites were chosen and *** present study focuses on the creation of fowlpox virus transfer vectors and its applications for multivalent vaccine vectors displaying multiple antigen ***:Based on the whole genome information of FPV282E4,FPV150(ortholog of vaccinia virus thymidylate kinase gene)and FPV193(ortholog of variola B22R-like protein)locus were chosen as insertion sites for foreign genes,and two shuttle vectors with a triple-gene expression cassette were designed and *** recombination occurred between the fowlpox virus genome and homologous sequences within the shuttle plasmids in infected *** recombinants were obtained through several rounds of plaque purification using enhanced green fluorescent protein as reporter genes and evaluated for the correct expression of foreign genes in vitro using RT-PCR,real-time PCR and Western *** and growth kinetics assays were tested via electron microscopy and viral titer ***:Two plasmid transfer vectors pT3eGFP150 and pT3eGFP193 were successfully constructed,Homologous recombination with fowlpox virus shuttle vectors pT3eGFP150 or pT3eGFP193 resulted in foreign exogenous genes inserting into the genomes of fowlpox virus,and caused that FPV 150 gene encoding was blocked or FPV 193 fragment

关键词： Fowlpox virus 282E4 strain Shuttle vectors Morphogenesis genetic stability

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Effect of TAV-free on physiology of Chrysanthemum morifolium Ramat.'Huaibai'

Effect of TAV-free on physiology of Chrysanthemum morifolium...

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中国植物病理学会2015年学术年会

作者：赵喜亭 TIAN Ying ZHU Yu-ting JIANG Li-Wei LI Ming-jun College of life science Henan normal university Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs of Henan Province University Henan Province Engineering Laboratory of Green Medicinal Plant Biotechnology

To explore the effect of tomato aspermyvirus(TAV)-free on physiology of Chrysanthemum morifolium Ramat.’ Huaibai’.TAV-free ’ Huaibai’ test-tube plantlets were used as experimental materials in this *** results showed that;(1) TAV-free didn’ t change the trend of photosynthetic indexes(the chlorophyll a,chlorophyll b and total chlorophyll contents) and N metabolism related indexes(the NR activity,Proline and soluble protein contents) of during subculture,but significantly increased the chlorophyll a contents,chlorophyll b contents and total chlorophyll contents at the same time,significantly increased the NR activity,Proline and soluble protein contents,so TAV-free improved photosynthetic capacity and N metabolism level.(2) TAV-free didn’ t change the trend of lipid peroxidation related indexes(O · produce rate,the MDA and HO contents) and protection system related indexes(protective enzymes SOD,POD,CAT,GR and APX activities;antioxidants ASA and GSH contents) during subculture,but significantly reduced the absolute value of lipid peroxidation related indexes and significantly increased activities of protective enzyme and the contents of antioxidants at the same time of *** TAV-free reduced the active oxygen metabolism and improved its oxidation resistance.

关键词： Chrysanthemum morifolium Ramat.’Huaibai’ Tomato aspermy virus virus-free genetic stability physiology

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