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Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

Progress in Natural Science:Materials International 2008年第7期18卷 819-824页

作者： Junfang Lin a,b, Mingyao Zheng a, Jie Wang a,b, Wei Shu c, Liqiong Guo a,b,* a Department of biotechnology, College of Food Sciences, South China agricultural University, Guangzhou 510640, China b Institute of Biomass Research, South China agricultural University, Guangzhou 510640, China c Test and Experimental center, South China agricultural University, Guangzhou 510640, China Department of biotechnology College of Food Sciences South China Agricultural University Guangzhou 510640 China Institute of Biomass Research South China Agricultural University Guangzhou 510640 China Test and Experimental center South China Agricultural University Guangzhou 510640 China

An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleu- rotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a "reporter" gene, green fluorescent protein gene can be successfully stably expressed in this Pleurotus species.

关键词： Pleurotus nebrodensis Protoplast-mediated transformation Green fluorescent protein GFP expression

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Improving maize grain yield by formulating plant growth regulator strategies in North China

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Journal of Integrative Agriculture 2021年第2期20卷 622-632页

作者： GONG Li-sha QU Shu-jie HUANG Guan-min GUO Yu-ling ZHANG Ming-cai LI Zhao-hu ZHOU Yu-yi DUAN Liu-sheng Engineering Research center of Plant Growth Regulator Ministry of Education/College of Agronomy and BiotechnologyChina Agricultural UniversityBeijing 100193P.R.China Experimental Base Service center of agricultural Sciences Jiyang 251400P.R.China

Plant growth regulators(PGRs) are artificially synthesized compounds that have become an important technical guarantee for agricultural production. EDAH(containing 27% ethephon and 3% DA-6) has been proven to inhibit stalk elongation, promote stalk bold and increase mechanical strength and number of vascular bundles. DA-6 could enhance plant photosynthetic capacity and promote cell division and growth. In our study, experiments were performed at summer maize growing season during 2018–2019. The result showed that plant height, ear height and center of gravity height of maize with EDAH+DA-6 treatment were decreased by 10.18, 16.77 and 13.21%, respectively;leaf area and leaf area index also significantly(P<0.001) decreased by 24.11 and 60.15%, respectively;the value of mean tilt angle significantly(P<0.001) increased by 16.72% compared with the control plants, which meant that EDAH+DA-6 could shape more compact plant type. Therefore, lodging rate of maize with EDAH+DA-6 treatment decreased by 6.95% compared with control plants, and the grain yield was increased by 15.51%. In addition, EDAH+DA-6 treatment significantly improved the quality of maize base stalks, such as improving mechanical properties, which increased maize base stalk crushing strength by 22.23%;increased the hemicellulose, cellulose and lignin contents by 6.93, 3.87 and 30.21%, respectively. In conclusion, EDAH+DA-6 treatment could improve summer maize yield by shaping plant morphological characteristics and group photosynthesis.

关键词： plant grown regulators canopy structure lodging resistance photosynthetic

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Immunolocalization of gap junction proteins-like matter at mesophyll protoplasts from Vicia faba

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Chinese Science Bulletin 1996年第15期41卷 1284-1290页

作者：潘照明娄成后 Laboratory of Molecular Biology Biotechnology Research Center Chinese Academy of Agricultural Sciences Beijing 100081 China Department of Plant Physiology and Biochemicstry Beijing agricultural University Beijing 100094 China

Intercellular connections and communications play very important roles during organisms growth and development. Although plasmodesmata and gap junction as intercellular channels are different in anatomy and structure, they exhibit striking similarities in their functions. It is suggested that they have some similar biochemical regulation mechanism. Connexins are the transfer of cytoplasmic material by conformational changes.

关键词： immunolocalization protoplast gap junction plasmodesmata.

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Effects of cultivation of Osr HSA transgenic rice on functional diversity of microbial communities in the soil rhizosphere

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The Crop Journal 2015年第2期3卷 163-167页

作者： Xiaobing Zhang Xujing Wang Qiaoling Tang Ning Li Peilei Liu Yufeng Dong Weimin Pang Jiangtao Yang Zhixing Wang biotechnology Research Institute Chinese Academy of Agricultural Sciences Biology Institute Hebei Academy of Sciences Development center of Science and Technology Ministry of Agriculture

With the widespread cultivation of transgenic crops, there is increasing concern about unintended effects of these crops on soil environmental quality. In this study, we used the Biolog method and ELISA to evaluate the possible effects of Osr HSA transgenic rice on soil microbial utilization of carbon substrates under field conditions. There were no significant differences in average well-color development(AWCD) values, Shannon–Wiener diversity index(H), Simpson dominance indices(D) and Shannon–Wiener evenness indices(E) of microbial communities in rhizosphere soils at eight samplings between Osr HSA transgenic rice and its non-transgenic counterpart. The main carbon sources utilized by soil microbes were carbohydrates, carboxylic acids, amino acids and polymers. The types,capacities and patterns of carbon source utilization by microbial communities in rhizosphere soils were similar throughout the detection period. We detected no Osr HSA protein in the roots of Osr HSA transgenic rice. We concluded that Osr HSA transgenic rice and the r HSA protein it produced did not alter the functional diversity of microbial communities in the rhizosphere.

关键词： Biolog method ELISA Soil microbes Functional diversity

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Dietary sanguinarine supplementation recovers the decrease in muscle quality and nutrient composition induced by high-fat diets of grass carp(Ctenopharyngodon idella)

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Animal Nutrition 2024年第2期17卷 208-219页

作者： Yong Shi Lei Zhong Yuanxiang Liu Shude Xu Jihong Dai Yaozhengtai Zhang Yi Hu Fisheries College Hunan Agricultural University Hunan Engineering Research center for Utilization of Characteristics of Aquatic Resources Hunan Agricultural University Key Laboratory of Breeding biotechnology and Sustainable Aquaculture Institute of Hydrobiology Chinese Academy of Sciences

The intake of high-fat diets(HFD) has been shown to diminish the muscle quality of aquatic ***, as an excellent additive, exhibits the capability to reduce fat deposition and alleviate inflammation. However, its role in the muscle quality reduction caused by HFD remains unclear. An eight-week trial was conducted to investigate the impacts of dietary supplementation of sanguinarine at1200 μg/kg(HFDS; crude fat = 10%) on the muscle quality of grass carp(Ctenopharyngodon idellus) in comparison to a basic diet(CON, crude fat = 5%). Each group had 3 replicates, with 40 fish per *** experiment employed one-way ANOVA and Duncan's multiple comparisons of the means. The results showed that the HFD exhibited lower growth performance, reduced protein deposition, myofiber diameter, and muscle hardness, coupled with higher levels of fat deposition and inflammation when compared with the CON. However, HFDS improved growth performance(P < 0.05), fat metabolism(ppara( P = 0.001), lpl(P < 0.001), atgl(P < 0.001), and cpt1(P = 0.001) expression exhibited a significant elevation), protein deposition(the protein and m RNA levels of AKT(P = 0.004), PI3K(P = 0.027),TOR(P = 0.005), and P70S6K(P = 0.007) demonstrated a marked increase), myofiber diameter, muscle hardness, and the total content of eicosapentaenoic acid and docosahexaenoic acid. Furthermore, the HFDS reduced oxidative damage caused by fat deposition by significantly downregulating nf-κb(P <0.001), il-1β(P < 0.001), il-6(P < 0.001), il-8(P = 0.003), and tnf-a(P < 0.001) expression and markedly upregulated nrf2(P < 0.001), gpx4(P < 0.001), cat(P < 0.001), sod(P < 0.001), and gr(P = 0.003)expression. The findings from this study suggest that sanguinarine has the potential to alleviate the adverse effects of HFD on growth and muscle quality, providing a theoretical foundation for its practical implementation.

关键词： Plant extract Muscle feature Myofiber characteristics Muscular texture Protein metabolism

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Timing effect of high temperature exposure on the plasticity of internode and plant architecture in maize

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Journal of Integrative Agriculture 2024年第2期23卷 551-565页

作者： Binbin Li Xianmin Chen Tao Deng Xue Zhao Fang Li Bingchao Zhang Xin Wang Si Shen Shunli Zhou College of Agronomy and biotechnology China Agricultural UniversityBeijing 100193China China Tobacco Hunan Industrial Corporation Changsha 410019China Innovation center of agricultural Technology for Lowland Plain of Hebei Wuqiao 061802China

The occurrence of high temperature(HT)in crop production is becoming more frequent and unpredictable with global warming,severely threatening food *** state of an organ’s growth and development is largely determined by the temperature conditions it is exposed to over *** is the main cereal crop,and its stem growth and plant architecture are closely related to lodging resistance,and especially sensitive to ***,systematic research on the timing effect of HT on the sequentially developing internode and stem is currently *** identify the timing effect of HT on the morphology and plasticity of the stem in maize,two hybrids(Zhengdan 958(ZD958),Xianyu 335(XY335))characterized by distinct morphological traits in the stem were exposed to a 7-day HT treatment from the V6 to V17 stages(Vn presents the vegetative stage with n leaves fully expanded)in *** results demonstrated that exposure to HT during V6-V12 accelerated the rapid elongation of *** instance,HT occurring at V7 and V12 specifically promoted the lengths and weights of the 3rd-5th and 9th-11th internodes,***,HT slowed the growth of internodes adjacent to the promoted ***,compared with control,the plant height was significantly increased soon after HT treatment,but the promotion effect became narrower at the subsequent flowering stage,demonstrating a self-adjusting mechanism in the maize plant in response to ***,HT altered the plant architectures,including a rising of the ear position and increase in the ear position ***335 exhibited greater sensitivity in stem development than ZD958 under HT *** findings improve our systematic understanding of the plasticity of internode and plant architecture in response to the timing of HT exposure.

关键词： maize high temperature internode growth plasticity plant architecture

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The Potential Efficacy of Glycyrrhizic Acid and Its Nanostructure Against Brown Rot of Peach fruits

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Phyton-International Journal of Experimental Botany 2023年第4期92卷 1139-1152页

作者： Tahsin Shoala Basma H.Amin Ismail A.S.Rashid Fayz A.Abdel-Rahman Mohamed E.Khalil Khamis Youssef Environmental biotechnology Department College of BiotechnologyMisr University for Science and Technology6th of October CityEgypt The Regional center for Mycology and biotechnology(RCMB) Al-Azhar UniversityCairo11435Egypt Plant Pathology Research Institute Agricultural Research CenterGiza12619Egypt

Production of peaches(Prunus persica(L.)Batsch)for both local market and export is increasing each year in *** rot disease,caused by Monilinia laxa and Monilinia fructigena,is considered one of the most important postharvest rots affecting peaches in Egypt and economic losses are *** activity of glycyrrhizic acid nanoparticles(GA-NPs)and glycyrrhizic acid(GA)at 0.2 and 0.4 mmol/L was investigated as a control for both these brown rot pathogens on peach fruits in both in vitro and in vivo *** the in vitro studies,GA-NPs were the most effective as shown by the ability to decrease linear growth of both brown rot pathogens in potato dextrose agar(PDA)amended with 0.4 mmol/L *** of *** exposed to 0.4 mmol/LGA showed mycelial deformations,nodule formation,detachment of the cell wall,shrinkage and inhomogeneous cytoplasmic materials with large *** of *** exposed to 0.4 mmol/LGA-NPs resulted in thinner and distorted hyphae,nodule formation,cell wall thinning,and *** GANPs and GA treatments improved fruit quality by maintaining firmness and total soluble solids(TSS).GA-NPs were more effective in decreasing decay incidence than their bulk *** 0.4 mmol/L GA-NPs completely inhibited the disease on naturally infected peach fruits for both seasons of 2018 and ***,0.4 mmol/L GA-NPs reduced the disease incidence in inoculated fruits by 95(***)and 88%(***)in 2018 season and 96(***)and 85%(***)in 2019 *** conclusion,GA-NPs could enhance the resistance of peaches against brown rot caused by *** and ***.

关键词： Prunus persica Monilinia laxa Monilinia fructigena glycyrrhizic acid peach postharvest

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Calcium contributes to photoprotection and repair of photosystem II in peanut leaves during heat and high irradiance

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Journal of Integrative Plant Biology 2015年第5期57卷 486-495页

作者： Sha Yang Fang Wang Feng Guo Jing-Jing Meng Xin-Guo Li Shu-Bo Wan biotechnology Research center Shandong Academy of Agricultural Sciences Shandong Provincial Key Laboratory of Crop Genetic Improvement Ecology and Physiology College of Life Sciences State Key Laboratory of Crop BiologyShandong Agricultural University

In this study, we investigated the effects of exogenous calcium nitrate on photoinhibition and thylakoid protein level in peanut plants under heat （40 ℃） and high irradiance （HI）（1,200 mmol/m2 per s） stress. Compared with control seedlings （cultivated in 0 mmol/L Ca（NO3）2 medium）, the maximal photochemical efficiency of photosystem II （PSII） in Ca2t‐treated plants showed a slight decrease after 5 h stress, accompanied by lower degree of PSII closure （1‐qP）, higher non‐photochemical quenching, and lower level of membrane damage. Ca2t inhibitors were used to analyze the varieties of antioxidant enzymes activity and PSII proteins. These results indicated that Ca2t could protect the subunits of PSII reaction centers from photoinhibition by reducing the generation of reactive oxygen species. In the presence of both ethyleneglycol‐bis（2‐aminoethylether）‐tetraacetic acid and ascorbic acid （AsA）, the net degradation of the damaged D1 protein was faster than that only treated with AsA. Our previous study showed that either the transcriptional or the translational level of calmodulin was obviously higher in Ca2t‐treated plants. These results suggested that, under＆amp;nbsp;heat and HI stress, the Ca2t signal transduction pathway can al eviate the photoinhibition through regulating the protein repair process besides an enhanced capacity for scavenging reactive oxygen species.

关键词： Calcium D1 protein heat and high irradiance stress peanut photosystem II

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Identification and Evaluation of Insect and Disease Resistance in Transgenic Cry1Ab13-1 and NPR1 Maize

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Phyton-International Journal of Experimental Botany 2023年第4期92卷 1257-1274页

作者： Yongjing Xi Zhou Yang Yukun Jin Jing Qu Shuyan Guan Siyan Liu Piwu Wang Joint Laboratory of International Cooperation in Modern Agriculture Technology of Ministry of Education Plant Biotechnology CenterJilin Agricultural UniversityChangchunChina

PCR detection,quantitative real-time PCR(q-RTPCR),outdoor insect resistance,and disease resistance identification were carried out for the detection of genetic stability and disease resistance through generations(T2,T3,and T4)in transgenic maize germplasms(S3002 and 349)containing the bivalent genes(insect resistance gene Cry1Ab13-1 and disease resistance gene NPR1)and their corresponding wild *** indicated that the target genes Cry1Ab13-1 and NPR1 were successfully transferred into both germplasms through tested generations;q-PCR confirmed the expression of Cry1Ab13-1 and NPR1 genes in roots,stems,and leaves of tested maize *** addition,S3002 and 349 bivalent gene-transformed lines exhibited resistance to large leaf spots and corn borer in the field evaluation compared to the wild *** study confirmed that Cry1Ab13-1 and NPR1 bivalent genes enhanced the resistance against maize borer and large leaf spot disease and can stably *** findings could be exploited for improving other cultivated maize varieties.

关键词： Maize NPR1 gene Cry1Ab13-1 gene disease resistance insect resistance

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Synthesis and Degradation of the Major Allergens in Developing and Germinating Soybean Seed

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Journal of Integrative Plant Biology 2012年第1期54卷 4-14页

作者： Yong-Mei Wu Rong-Xia Guan Zhang-Xiong Liu Run-Zhi Li Ru-Zhen Chang Li-Juan Qiu center of agricultural biotechnology Shanxi Agricultural University Taigu 030801 China National Key Facility of Gene Resources and Genetic Improvement/Key Laboratory of Crop Germplasm Utilization the Ministryof Agriculture Institute of Crop Science Chinese Academy of Agricultural Sciences Beijing 100081 China

Gly m Bd 28K, Gly m Bd 30K and Gly m Bd 60K are the major soybean （Glycine max （L.） Merr.） allergens limiting the consumption of a good protein source for sensitive individuals. However, little is known about their temporal-spatial expression during seed development and upon germination. The present data shows that soy allergens accumulated in both the embryonic axes and cotyledon, but expression patterns differed depending on the specific allergen. Allergens accumulated sooner and to a greater level in cotyledons than in embryonic axes. Gly m Bd 28 began at 14 d after flowering, 7 to 14 d earlier than Gly m Bd 30K and Gly m Bd 60K. Comparatively, their degradation was faster and more profound in embryonic axes than in cotyledons. Gly m Bd 60K began to decline at 36 h after imbibition and remained detectable up to 108 h in cotyledons. In contrast, the Glym Bd 60K protein was reduced at 24 h, and eventually disappeared at 96 h. In cotyledons Gly m Bd 28K first declined at 24 h, then increased from 36 h to 48 h, followed by its large reduction at 72 h after seed germination. These findings provide useful information on soy allergen biosynthesis and will help move forward towards developing a hypoallergenic soybean for safer food.

关键词： allergen food allergy Glycine max soybean synthesis and degradation seed development and germination

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