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Different genotypes regulate the microbial community structure in the soybean rhizosphere
QU Zheng, LI Yue-han, XU Wei-hui, CHEN Wen-jing, HU Yun-long, WANG Zhi-gang
2023, 22 (2): 585-597.   DOI: 10.1016/j.jia.2022.08.010
Abstract244)      PDF in ScienceDirect      

The soybean rhizosphere has a specific microbial community, but the differences in microbial community structure between different soybean genotypes have not been explained.  The present study analyzed the structure of the rhizosphere microbial community in three soybean genotypes.  Differences in rhizosphere microbial communities between different soybean genotypes were verified using diversity testing and community composition, and each genotype had a specific rhizosphere microbial community composition.  Co-occurrence network analysis found that different genotype plant hosts had different rhizosphere microbial networks.  The relationship between rhizobia and rhizosphere microorganisms in the network also exhibited significant differences between different genotype plant hosts.  The ecological function prediction found that different genotypes of soybean recruited the specific rhizosphere microbial community.  These results demonstrated that soybean genotype regulated rhizosphere microbial community structure differences.  The study provides a reference and theoretical support for developing soybean microbial inoculum in the future.

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PbPH5, an H+ P-ATPase on the tonoplast, is related to malic acid accumulation in pear fruit
SONG Jun-xing, CHEN Ying-can, LU Zhao-hui, ZHAO Guang-ping, WANG Xiao-li, ZHAI Rui, WANG Zhi-gang, YANG Cheng-quan, XU Ling-fei
2022, 21 (6): 1645-1657.   DOI: 10.1016/S2095-3119(21)63790-5
Abstract203)      PDF in ScienceDirect      

Organic acids are one of the most important factors influencing fruit flavors. The predominant organic acid in most pear cultivars is malic acid, but the mechanism controlling its accumulation remains unclear. In this study, by comparing gene expression levels and organic acid content, we revealed that the expression of PbPH5, which encodes a P3A-ATPase, is highly correlated with malic acid accumulation in different pear species, with correlation coefficients of 0.932**, 0.656*, 0.900**, and 0.518* (*, P<0.05 or **, P<0.01) in Pyrus bretschneideri Rehd., P. communis Linn., P. pyrifolia Nakai., and P. ussuriensis Maxim., respectively. Moreover, the overexpression of PbPH5 in pear significantly increased the malic acid content. In contrast, silencing PbPH5 via RNA interference significantly decreased its transcript level and the pear fruit malic acid content. A subcellular localization analysis indicated that PbPH5 is located in the tonoplast. Additionally, a phylogenetic analysis proved that PbPH5 is a PH5 homolog gene that is clustered with Petunia hybrida, Malus domestica, and Citrus reticulata genes. Considered together, these findings suggest PbPH5 is a functionally conserved gene. Furthermore, the accumulation of malic acid in pear fruits is at least partly related to the changes in PbPH5 transcription levels.

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Exploration of the key microbes and composition stability of microbial consortium GF-20 with efficiently decomposes corn stover at low temperatures
YU Xiao-fang, BORJIGIN Qinggeer, GAO Ju-lin, WANG Zhi-gang, HU Shu-ping, BORJIGIN Naoganchaolu, WANG Zhen, SUN Ji-ying, HAN Sheng-cai
2019, 18 (8): 1893-1904.   DOI: 10.1016/S2095-3119(19)62609-2
Abstract169)      PDF in ScienceDirect      
The microbial consortium GF-20 (GF-20) can efficiently decompose corn stover at low temperatures.  The present study explored the key microbes of GF-20 and evaluated different culture conditions on its composition stability to promote the utilization of corn stover decomposing microbes in low temperature regions.  GF-20 was subcultured to the 15th generation under different temperatures, pHs, carbon, and nitrogen sources.  Then, the dynamics of fermenting pH, cellulose enzyme activities, carbohydrate concentration, and oxidation reduction potential were determined to estimate the degradation efficiency of corn stover with GF-20.  Furthermore, the structural stability and functional microbes of GF-20 were identified on the basis of PCR-denaturing gradient gel electrophoresis (DGGE) profiling and principal component analysis.  The results showed that the offspring of GF-20 subcultured under different temperatures (4–30°C) and pH (6.0–9.0) conditions maintained stable growth, decomposition function, and composition structure.  Furthermore, consortia GF-20 had a stable composition structure, which induced GF-20 to secrete cellulose and promote substrate decomposition as corn stover and ammonium were used as sources of carbon and nitrogen, respectively.  According to the PCR-DGGE profiles, the key strains of GF-20 were determined to be Bacillus licheniformis, Cellvibrio mixtus subsp. mixtus, Bacillus tequilensis, Clostridium populeti, and Clostridium xylanolyticum.
 
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Screening of a microbial consortium with efficient corn stover degradation ability at low temperature
Qinggeer , GAO Ju-lin, YU Xiao-fang, ZHANG Bao-lin, WANG Zhi-gang, Borjigin Naoganchaolu, HU Shu-ping, SUN Ji-ying, XIE Min, WANG Zhen
2016, 15 (10): 2369-2379.   DOI: 10.1016/S2095-3119(15)61272-2
Abstract1442)           
    To speed up the degradation of corn stover directly returned to soil at low temperature, the corn stover-degrading microbial consortium GF-20, acclimated to biological decomposition in the frigid region, was successfully constructed under a long-term limiting substrate. To evaluate its potential in accelerating the decomposition of un-pretreated corn stover, the decomposing property, fermentation dynamic and the microbial diversity were analyzed. GF-20 degraded corn stover by 32% after 15-day fermentation at 10°C. Peak activities of filter paperlyase (FPA), β-glucosidases (CB), endoglucanases (Cx), and cellobiohydrolases (C1) were 1.15, 1.67, 1.73, and 1.42 U mL–1, appearing at the 6th, 3rd, 11th, and 9th d, respectively. The pH averaged at 6.73–8.42, and the optical density (OD) value peaked at 1.87 at the 120 h of the degradation process. Cellulase, hemicellulase and lignin in corn stover were persistently degraded by 44.85, 43.85 and 25.29% at the end of incubation. Result of denaturing gradient gel electrophoresis (DGGE) profiles demonstrated that GF-20 had a stable component structure under switching the temperature and pH. The composition of the GF-20 was also analyzed by constructing bacterial 16S rDNA clone library and fungal 18SrDNA-PCR-DGGE. Twenty-two bacterial clones and four fungal bands were detected and identified dominant bacteria represented by Cellvibrio mixtus subsp., Azospira oryzae, Arcobacter defluyii, and Clostridium populeti and the fungi were mainly identified as related to Trichosporon sp.
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Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation
ZHAO Xi-an, CANG Ming, GAO Xiao-yu, YANG Mei-ling, YUAN Jian-long, ZHU Bing, WANG Zhi-gang
2012, 12 (8): 1333-1339.   DOI: 10.1016/S1671-2927(00)8663
Abstract1422)      PDF in ScienceDirect      
To study the effects of interleukin-6 (IL-6) and its receptor (IL-6R) during in vitro maturation of ovine oocytes, the mRNA and protein expression levels of IL-6 and IL-6R, along with their localization, were examined during ovine oocytes maturation in vitro through real-time PCR, Western blotting, and immunohistochemistry. Specific patterns of expression of IL-6 and IL-6R were observed at both mRNA and protein levels at each stage of ovine oocytes maturation. IL-6 and IL-6R were distributed primarily on the surface of the cell membrane, with little expression in the cytoplasm or nucleus. IL-6 and IL- 6R were expressed significantly at higher levels in the maturation around 4 h, and then decreased dramatically. However the level slightly elevated at 20-24 h. The role of IL-6 and IL-6R on oocytes maturation was studied through in vitro addition of recombinant human IL-6 in different concentrations. The addition of 10 ng mL-1 IL-6 significantly increased the rates of oocytes maturation (P<0.05), but did not affect the rates of development of the subsequence IVF ovine embryos. In summary, IL-6 is likely to play an important role in the early ovine oocytes maturation. The expression patterns of the IL-6 and IL-6R on the ovine oocytes maturation open up the possibility of regulatory role of the cytokine in ovine oocytes maturation.
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Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus) 
ZHENG Xu, YANG Jiao-fu, WANG Xiao-jing, LIANG Yan, WU Man-lin, SHI Jie-jun, ZHANG Tao, QIN Yin, LI Shu-yu, HAO Xi-yan, WANG Zhi-gang , LIU Dong-jun
2011, 10 (9): 1452-1458.   DOI: 10.1016/S1671-2927(11)60138-7
Abstract1565)      PDF in ScienceDirect      
As one member of the Ras super family, Rheb is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, proliferation and differentiation. In order to study the relationship between Rheb and mTOR in Inner Mongolian Cashmere goat (Capra hircus) cells, Ras homolog enriched in brain (Rheb) gene cDNA was amplified by RT-PCR. It is 555 bp in length and includes the complete ORF encoding 184 amino acids (GenBank accession no. HM569224). The full cDNA nucleotide sequence has a 99% identity with that of sheep, 98% with cattle and 93% with human while their amino acids sequence shares identity with 98, 97 and 97% of them, correspondingly. The bioinformatics analysis showed that Rheb has a Ras family domain, two casein kinase II phosphorylation sites, two ATP/GTP-binding sites motif A (P-loop), a prenyl group binding site (CAAX box). Tissue-specific expression analysis performed by semiquantitative RT-PCR. The Rheb gene was expressed in all the tested tissues and the highest level of mRNA accumulation was detected in brain, suggesting that Rheb played an important role in goat cells.
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