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Microbial Diversity in Rhizosphere and Non-rhizosphere Soils of Paris polyphylla var. chinensis Plants
ZHENG Meixia, CHEN Hong, ZHU Yujing, SU Hailan
 doi: 10.19303/j.issn.1008-0384.2020.12.012
[Abstract](122) [FullText HTML](57) [PDF 1146KB](4)
Abstract:
  Objective   The microbial communities in rhizosphere and non-rhizosphere soils growing Paris polyphylla var. chinensis plants were analyzed and compared in the field as well as in laboratory.   Method   The high throughput sequencing Illumina Miseq 2×300 bp was applied to sequence total DNA of the microbes in the soils. The LDA Effect Size analysis was used to examine the differences on selected microbial groups between the communities in the field and laboratory.   Result   The microbial diversity in the soils of P. polyphylla was rich with 21 phyla, 33 classes, 52 orders, 89 families, and 160 genera identified. The Chao, Ace, Shannon, and Simpson indices on the soil samples indicated that the non-rhizosphere soil was more abundant and diverse than the rhizosphere soil. However, there were no significant differences between them on the phylum or family level. Among the differentially abundant groups, Chlorobia on the class level, Chlorobiales and Myxococcales on the order level, and Acinetobacter and Rudaea on the genus level were significantly lower in the rhizosphere.   Conclusion  This study not only shows that Paris polyphylla var. chinensis non-rhizosphere soil bacterial communities have higher abundance and diversity, but also lays the foundation for further exploration of the soil microbial environment of Paris polyphylla var. chinensis.
Construction of a yeast two-hybrid cDNA library from Vitis davidii Föex pericarp infected by grape ripe rot (Colletotrichum viniferum)
LEI Yan, XIE Qian, CHEN Ting, LIU Xin-ming, CHEN Qing-xi
[Abstract](119) [FullText HTML](67) [PDF 1171KB](19)
Abstract:
  Objective  Grape ripe rot is an important disease in southern China. The spine grape (Vitis davidii Föex), an important wild grape species, is highly resistant to grape ripe rot. At present, there is no report about the yeast two-hybrid cDNA library of grape pericarp after grape ripe rot (Colletotrichum viniferum) infection. In this study, a yeast two-hybrid cDNA library of grape pericarp after grape ripe rot (C. viniferum) infection was constructed to provide a reference for resistant molecular mechanism of C. viniferum.  Method  Total RNA was extracted from the pericarp of V. davidii accession 'Fu'an' at 1 d, 3 d and 7 d after infected by C. viniferum. Total RNA was reverse transcription into cDNA by The SMART method. The cDNA was purified by using the kit assay, and then the purified product (double stranded cDNA) was digested, then the short fragments were removed to obtain the high-quality cDNA. The cDNA was cloned into pGADT7 three frames plasmid vector and purified to obtain the primary cDNA library. After the primary cDNA library was amplified, the plasmid was extracted and transformed into yeast Y187 to produce the amplified yeast library. Finally, the yeast two-hybrid cDNA library was identified.  Result  The results showed that the capacity of primary cDNA library was about 5.2 ×106 cfu, the recombination rate was about 97.92%, and the length of inserted fragments ranged from 400 to 2000 bp with good polymorphism, harvested Y187 yeast library titer was about 6.0×107 cfu·mL−1.  Conclusion  A yeast two-hybrid cDNA library of grape pericarp after grape ripe rot infection was constructed in this study and it qualified for yeast two-hybrid screening. This yeast two-hybrid cDNA library provided a reference to screen the interaction proteins in grapes under grape ripe rot infection.
Identification of the blast resistance genes via BSA-Seq method in rice cultivar Gufeng B
CHEN Ziqiang, CHEN Songbiao, GUO Xinrui, YAN Jingwan, TIAN Dagang, LI Gang, WANG Feng
[Abstract](88) [FullText HTML](60) [PDF 500KB](2)
Abstract:
  Objective  The rice cultivar GuFeng B confers strong, broad-spectrum, durable resistance against various rice blast isolates. The present study was aim to identify and map the blast resistance gene(s) in Gufeng B.  Method  The F1 and F2 population were obtained by crossing Gufeng B and Nipponbare, and the genetic model of blast resistance was analyzed after inoculated with Magnaporthe grisea 501-3. Subsequently, F2 population was used to construct a resistant pool and a sensitive pool respectively, and to map the associated loci via the method of bulked segregation analysis.  Result  Gufeng B exhibited high resistance to all of the tested strains, such as KJ201, RB22, CHNOS, RB6, 2Y838-1, 501-3 and IR16-1, suggesting that Gufeng B may carry the broad-spectrum and high resistance genes. The F1 progenies from the cross between Gufeng B and Nipponbare conferred resistance against the strains 501-3 and IR16-1, and the segregation ratio of resistance and susceptibility among F2 progenies does not fit 3:1, assuming that the resistance against the strains 501-3 and IR16-1 were controlled by multiple locus in Gufeng B. Whole genome re-sequencing of the two parental lines Gufeng B and Nipponbare identified 1,756,964 SNPs. Calculation results of △SNP-index showed that there were two candidate loci conferring resistance to rice blast disease, which were located at Chr.6: 10,082-11,397Kb, corresponding to the Pi2/9 locus, and Chr.11: 120-266Kb. 4006 SNPs and 623 InDels markers were searched within the interval of Chromosome 6, 752 SNPs and 195 InDels within the corresponding region of Chromosome 11, respectively.  Conclusion  The resistance of Gufeng B to 501-3 strain may be controlled by two resistance genes on chromosomes 6 and 11. Our results laid the foundation for finely mapping and cloning the resistance genes in Gufeng B, and provided marker resources for molecular marker-assisted selection.
Transcriptome analysis of root tubers of two cultivated species of Tetrastigma hemsleyanum Diels et Gilg in Huaiyushan
HONG Senrong, HUANG Dandan, HUANG Shihui, HUANG Xiameng, JIANG Ye, LI Wanping, CAI Hong, CHEN Ronghua
[Abstract](96) [FullText HTML](46) [PDF 1250KB](1)
Abstract:
  Objective  Screening of differentially expressed genes related to flavonoid synthesis in two cultivated species (‘Huaiyu1’ and ‘Huaiyu2’) of Tetrastigma hemsleyanum Diels et Gilg in Huaiyushan.  Method  This study used the root tubers of ‘Huaiyu1’ (Group HY1) and ‘Huaiyu2’ (Group HY2) as experimental materials for transcriptome analysis.  Result  The clean reads of group HY1 and group HY2 were 42311662 and 41411202, respectively. The percentage of Q30 base in 2 groups was not less than 95.75%. The transcription factor families of HY1 and HY2 were MYB-superfamily, bHLH, AP2/ERF, NAC, C2C2, WRKY, etc. The paired value of FPKM in HY1 group and HY2 group was between 0-2. The expression density of HY1 group and HY2 group was between 0-0.7. The number of common genes expressed in HY1 and HY2 groups was 22367, the number of genes separately expressed in group HY1 was 18196, the number of genes independently expressed in HY2 group was 8137. The correlation coefficient of HY1 group and HY2 group was 0.913, and the correlation between samples was good. There were 12199 differentially expressed genes in HY1 and HY2 groups. Compared with HY1 group, the number of up-regulated genes and down-regulated genes in HY2 group were 3551 and 8648 respectively. GO Enrichment analysis showed that the differential genes were mainly annotated into photosynthesis, light harvesting in photosystem I, photosynthesis, light harvesting, chlorophyll metabolic process, protein-chromophore linkage, generation of precursor metabolites and energy, chlorophyll biosynthetic process, response to oxidative stress, alpha-amino acid metabolic process, photosynthesis, plastoglobule, photosystem I, photosystem II, plastid nucleoid, photosystem, chlorophyll binding, monooxygenase activity, iron ion binding, heme binding, lyase activity, etc. KEGG Enrichment analysis showed that the differential genes were mainly annotated to photosynthesis-antenna proteins, ribosome, glyoxylate and dicarboxylate metabolism, phenylpropanoid biosynthesis, stilbenoid, diarylheptanoid and gingerol biosynthesis, flavonoid biosynthesis, photosynthesis, carbon fixation in photosynthetic organisms, glycine, serine and threonine metabolism, plant hormone signal transduction, glutathione metabolism, pyruvate metabolism, phenylalanine metabolism, circadian rhythm-plant, flavone and flavonol biosynthesis, cysteine and methionine metabolism, cyanoamino acid metabolism, carotenoid biosynthesis, alpha-linolenic acid metabolism, porphyrin and chlorophyll metabolism, and other metabolic pathways.  Conclusion  The differentially expressed genes related to flavonoids, such as stilbene synthase, leucoanthocyanidin dioxygenase, Chi protein, chalcone synthase 2, flavanone 3-hydroxylase and lleucoanthocyanidin reductase 1 and flavonoid 3′-hydroxylase gene were up-regulated in ‘Huaiyu 2’ (HY2 group), while chalcone synthase, flavonol synthase and flavonoid 3′, 5′- methyltransferase were down-regulated in root tubers of' ‘Huaiyu 2’ (HY2 group), which resulted in the difference of total flavonoids content between ‘Huaiyu 1’ (HY1 group) and ‘Huaiyu 2’ (HY2 group).
Response characteristics of endangered semi mangrove plant Barringtonia racemosa to tidal flooding
LIANG Fang, PAN Yanju, DENG Xu, WU Yushuang, LIANG Zerui, ZHAO Shihua, TAN Xiaohui
[Abstract](93) [FullText HTML](64) [PDF 1171KB](2)
Abstract:
  Objective  To analyze the response characteristics of the endangered semi-mangrove plant Barringtonia racemosa when it is submerged by tides, to provide a scientific basis for the selection of suitable flooding time for B. racemosa in urban inland rivers, lakesides, wetland restoration and other landscape applications.  Method  The two-year-old seedlings of B. racemosa seedlings were used as materials to simulate semi-diurnal tides to study the plant morphology, physiology, biochemistry and response characteristics of mineral elements when the seedlings were submerged by tides with different flooding durations.  Results  (1) B. racemosa seedlings can survive 250 days of tidal submergence stress treatment, and their plant height, number of leaves, and leaf area are significantly lower than those of control plants. The numbers were significantly higher than the control group. (2) Chlorophyll a and total chlorophyll of B. racemosa leaves (except 16 h/d) showed an overall downward trend; MDA content showed an overall upward trend (except 14 h/d), and the content of proline and soluble sugar was submerged for 10 h/d. It reached the maximum value at d and was significantly higher than that of CK; (3) Flooding stress promoted the absorption of N, P, and Fe elements in the leaves, but inhibited the absorption of K and Cu elements.  Conclusion  Leaf area, leaf phosphorus content, total chlorophyll content, lateral root surface area, CAT activity, POD activity and soluble protein content were the main indexes of B. racemosa responding to flooding stress. B. racemosa has strong tolerance and adaptability to semi-diurnal tide flooding adversity with flooding time ≤20 h/d, and It can be planted in the waterfront, lakeshore green belt and freshwater wetlands and other landscape environments.
Screening of Microalgae for High Efficiency Removal of Nitrogen and Phosphorus from Aquaculture Wastewater
JIA Wei, NIE Yilei, CHEN Hong, LUO Lijin, LE Zhanxian, ZHUANG Hong, ZHENG Junrong
[Abstract](76) [FullText HTML](62) [PDF 666KB](0)
Abstract:
  Objective  It is aimed to screen high efficient functional microalgae on nitrogen and phosphorus removal in aquaculture wastewater.  Method  Efficiency of nitrogen and phosphorus removal from Penaeus vannamei mariculture wastewater by Chlorella sp.JY-1, Chlorella sp.SY-4 and Desmodesmus sp. SH-1 was studied.  Result  The results showed after 5 days of cultivation the cell density of three microalgae were 1.56×107 ml−1, 1.47×107 ml−1 and 6.62×106 ml−1 respectively. The removal rates of total nitrogen were 50.36%、41.51% and 49.74%. The removal rates of ammonia nitrogen were 96.29%, 84.92% and 96.65% respectively. The removal rates of nitrate nitrogen were15.84%, 3.69% and 12.56% respectively, and the removal rates of total phosphorus were 93.51%, 82.38% and 94.25% respectively. However, nitrite nitrogen concentration in the wastewater was not significantly removed. Three microalgae can grow normally in culture medium with 5%, 10%, 20% and 30% salinity. Compared with Chlorella sp.SY-4 and Desmodesmus sp. SH-1, Chlorella sp.JY-1 has better growth ability and efficiency of nitrification and phosphorus removal from aquaculture wastewater.  Conclusion  This study provides a theoretical basis for the application of microalgae in the purification of aquaculture wastewater.
Research progress on HD-ZIP transcription factor under pathogens and abiotic stress response
Liu jiao, Shuai peng
[Abstract](158) [FullText HTML](95) [PDF 953KB](1)
Abstract:
Adverse environmental conditions usually affect plant growth and development, indirectly or directly causing crop production reduction and even plant death. HD-ZIP transcription factors have been described to be involved in stress responses.The HD-ZIP transcription factor, which belongs to the homeobox (HB) protein family, is unique to plants that are tightly connected by the highly conserved homeobox (HD) and leucine zipper domain (ZIP).The formation of protein dimers mediated by the LZ domain allows the HD domain to bind to the target DNA and regulate the expression of the target gene. HD-ZIP transcription factors not only play an important role in regulating plant growth and development, but are also critical for responding to adverse environmental stress. This review focuses on the role of the four subfamilies (Ⅰ-Ⅳ) of HD-ZIP transcription factors responses under various pathogens and abiotic stress, such as drought, salt, extreme temperature, wounding, low R/FR light and heavy metal stress based on research reports in recent years. Here we discuss the internal molecular mechanism of HD-ZIP transcription factors on how to integrate hormones and environmental signals to improve plant growth characteristics, thus laying a foundation for plants to improve stress resistance.
Cloning and Bioinformatics Analysis of Protein Phosphatase Genes MiSTPP1 and MiSTPP4 from Macadamia integrifolia
LI Jidong, YANG Xiangyan, CAI Yuanbao, LI Mu, ZENG Liming, HUANG Sijie, WU Fumin, LIN Yuhong, ZHENG Wenwu, HUANG Jinyuan
[Abstract](143) [FullText HTML](79) [PDF 960KB](0)
Abstract:
  Objective  Protein phosphatase genes MiSTPP1 and MiSTPP4 were cloned and analyzed the structure and function by bioinformatics, which provided the theoretical reference for the function and mechanism of protein phosphatases in macadamia (Macadamia integrifolia).  Method  Protein phosphatase genes were cloned from M. integrifolia by transcriptome sequencing and RT-PCR technique. The homology, phylogenetic evolution, physicochemical properties, phosphorylation sites, subcellular localization, transmembrane domains and signal peptides of protein phosphatases were analyzed by bioinformatics.  Result  Two new protein phosphatase genes MiSTPP1 and MiSTPP4 were cloned from M. integrifolia (GenBank accession numbers are MT374548 and MT374551). Amino acid homology analysis showed that MiSTPP1 and MiSTPP4 had high sequence similarity with other plant PP1 proteins, which contained the typical structural domain MPP_PP1_PPKL. Evolutionary tree analysis showed that MiSTPP1 and MiSTPP4 were closely related to PP1 family proteins. The analysis of the basic physicochemical properties of the protein showed that MiSTPP1 was an unstable hydrophilic protein, and MiSTPP4 was a stable hydrophilic protein. Phosphorylation sites analysis showed that the phosphorylations of MiSTPP1 and MiSTPP4 were dominated by serine and threonine. The prediction of subcellular localization, transmembrane domain and signal peptide indicated that MiSTPP1 and MiSTPP4 as the non-secretory and non-transmembrane proteins were most likely located in the cytoplasm. In addition, the secondary and 3d structures of MiSTPP1 and MiSTPP4 mainly contain α - helixs and random coils.  Conclusion  MiSTPP1 and MiSTPP4 belong to the protein phosphatase PP1 gene family and may play a role in response to stress, signal transduction and other physiological and biochemical processes.
Effect of endogenous hormone content and gene expression related to hormone synthesis on tuber expansion of Bikeqi yam
AO Lan-ji-ya, JI Xiang, SHAO Ying, ZHAO Lingmin, ZHANG Yan-fang, HUO Xiu-wen
[Abstract](561) [FullText HTML](446) [PDF 693KB](14)
Abstract:
  Objective  To investigate the changes of starch content, reducing sugar content and soluble total sugar content in tubers, the changes of endogenous hormone content in tubers, the morphological indexes, physiological indexes and the analysis of variance of endogenous hormones during the process of tuber expansion. And the correlation between these indicators and endogenous hormones, lay the foundation for further study of the physiological mechanism of yam growth and development and guiding the high yield and quality cultivation of yam.  Method  Six kinds of endogenous hormones such as ABA, GA3, IAA, JA, ZR and IPA were determined by enzyme-linked immunosorbent assay with different tuber lengths of Bikeqi yam. High performance liquid chromatography (HPLC) was used to determine the content of salicylic acid (SA), and to analyze the dynamic changes of source hormone content in different growth stages, and the correlation between morphological indicators of yam and endogenous hormones. The relationship between differentially expressed genes associated with endogenous hormones and endogenous hormones was explored for the effects of these factors on the expansion of yam tubers.  Result  The contents of IAA, ZR, ABA, JA and SA were positively correlated with the morphological indexes of yam tuber; the contents of GA3 and IPA were negatively correlated with the morphological indexes; the contents of IAA were positively correlated with the perimeter and diameter of yam tuber; the contents of GA3 were negatively correlated with the length of tuber; the genes correlated with IAA were negatively correlated with the contents of IAA  Conclusion  Endogenous hormones IAA, ZR, ABA, JA and SA promote the expansion of yam tuber; endogenous hormones GA3 and IPA inhibit the growth of yam tuber; endogenous hormones IAA promote the thickening of yam; endogenous hormones GA3 inhibit the elongation of yam; the down-regulation of IAA related genes can promote the synthesis of IAA, that is, regulate the content of IAA.
2020, 35(11): 1-2.  
[Abstract](0) [FullText HTML](0) [PDF 299KB](0)
Abstract:
Crop Science
Breeding and Application of New Two-line Special Rice Variety Ziliangyou 737
HUANG Tingxu, ZHENG Jianhua, YOU Qingru, DONG Ruixia, WANG Hongfei, FANG Xiantao, LIAO Falian, CHEN Chunxia, ZHOU Peng, TU Shihang, ZHENG Feiyan, DONG Lianfei
2020, 35(11): 1171-1178.   doi: 10.19303/j.issn.1008-0384.2020.11.001
[Abstract](0) [FullText HTML](1) [PDF 759KB](0)
Abstract:
  Objective  To fully exploit and utilize precious purple-black rice resources, strengthen the innovation of special rice resources, expand the planting area of purple-black special rice, and satisfy people's demand for healthy diet.  Method  A new two-line special rice variety Ziliangyou 737 was cross bred from Zi 392S, a indica glutinous Dual-purpose Genic Male Sterile (DGMS) line with purple-black seed coat developed by Rice Research Institute, Fujian Academy of Agricultural Sciences and Fuhui 737, a self-breeding purple glutinous restorer line in the spring of 2014. The comprehensive trait performances of Ziliangyou 737 were investigated in variety comparative test during 2014-2016 in Fujian, Yunnan, Jiangxi and Anhui. During 2017-2018, it was recommended to join in the regional trial and production trial of the special-purpose purple rice groups independently organized by Yunnan State-owned Capital Operation Jinding Hepu Agricultural Technology Co., Ltd. And during 2018-2019, it was recommended to join in the regional trials of late-cropping special rice groups in Fujian in order to evaluate its comprehensive traits.  Result  The average yields of Ziliangyou 737 during 2014-2016 in Fujian, Yunnan, Jiangxi, Anhui and other local product comparative tests were more than 7 500.0 kg·hm−2; In the regional trial of special-purpose purple rice group in Yunnan in 2017-2018, the growth period was about 161 days, the plant height was 97.0 cm, the effective panicles were 191, 000 per 667 m2, the ear length was 22.9 cm, the total grains per ear were 179.9, the seed setting rate was 72.1%, 1000-grain weight was 24.9 g, the amylose content was 2.6%, the average yields of the regional trials were 7 564.2 kg·hm−2 and 7 569.3 kg·hm−2 respectively, increased by 148.0% and 156.2% compared to control Guineng purple rice; the average yields of the 2018 production trial was 7 964.7 kg·hm−2, increased by 169.2% compared to control Guineng purple rice; it was approved by the Yunnan Crop Variety Appraisal Committee in 2019 (Dianshendao 2019004). In the regional trials of the late rice special rice group in Fujian Province in 2018 and 2019, the growth period was about 123 days, the effective panicles were 163 000 per 667 m2, the plant height was 108.4 cm, the panicle length was 23.7 cm, and grain number per panicle was 172.4, the seed setting rate was 79.33%, 1 000-grain weight was 24.7 g, amylose content was 2.1%, the regional average yields was 7 475.6 kg/hm2 and 7,535.4 kg/hm2 respectively, decreased by 4.39% and 3.26% compared to control Yiyou 673, it was approved by the Fujian Crop Variety Appraisal Committee in 2020 (Minshendao 20200067).  Conclusion  Ziliangyou 737 has promising application prospects, with excellent comprehensive agronomic traits, strong heterosis, high yield stability, wide adaptability, deep purple brown rice seed coat, waxy endosperm, excellent rice quality, especially good food taste and other characteristics. It can be promoted as special rice in the hybrid indica rice area below 1 450 meters above sea level in Fujian and Yunnan provinces and other similar ecological regions.
Horticultural Science
Reference Gene Selection for RT-qPCR Analysis on Cucumis melo
ZHANG Lizhen, HAN Xiaoyun, WU Jinghua, Gefu WANG-PRUSKI, ZHANG Zhizhong
2020, 35(11): 1179-1187.   doi: 10.19303/j.issn.1008-0384.2020.11.002
[Abstract](93) [FullText HTML](123) [PDF 921KB](2)
Abstract:
  Objective  In search for internal reference genes of Cucumis melo L. that could stably express in different tissues and under stress conditions to warrant accuracy and reliability of the RT-qPCR analysis on target gene expression.   Method   Expression stabilities of 9 candidate genes, 18srRNA, TUA, EF1a, Actin1, Actin2, Actin3, Actin4, CYC, and UBI-ep, from the roots, leaves, seeds, and fruits of Xinyinhui melon being treated by water, cinnamic acid, saline alkali or ABA were determined by RT-qPCR and analyzed using the BestKeeper, NormFinder, and geNorm software.  Result   In different tissues, the top 5 choice genes identified by BestKeeper ranked as CYC18s rRNAUBI-epEF1aTUA, those by NormFinder EF1aUBIepActin4CYCActin3, and those by geNorm Actin4=Actin3Actin1EF1aUBI-ep. Under various stresses, they were 18s rRNAActin3Actin4EF1aUBI-ep as ranked by BestKeeper, EF1aUBI-epActin4CYC18s rRNA by NormFinder, and EF1a=UBI-epActin4CYCActin by geNorm. Overall, EF1a appeared to be most stable among the 9 genes. Insofar as variety of tissues is concerned, Actin4, Actin3, Actin1, and EF1a were more stable than the others; and, under stress, EF1a and UBI-ep tended to be superior.   Conclusion   Stably expressed in the tissues under the stresses as tested, EF1a was selected as the reference gene for studies on C. melo to reduce experimental errors. To further ensure accuracy, application of dual reference genes in RT-qPCR analysis might be considered.
Genome-wide Identification and Expression Analysis of Metacaspase Gene Family in strawberry
ZHENG Yiping, CHEN Jian, ZHU Bingyao
2020, 35(11): 1188-1197.   doi: 10.19303/j.issn.1008-0384.2020.11.003
[Abstract](85) [FullText HTML](41) [PDF 1148KB](3)
Abstract:
  Objective  To identify and study the genes in strawberry (Fragaria ananassa) which related to the synthesis of metacaspases (MCs), the enzymes that played vital roles in regulating the programmed cell death (PCD) in plants.   Method  The physiochemical properties, conserved domains, phylogenetic relationships and conserved motifs of FaMC proteins were analyzed based on strawberry whole genome data. The expressions of FaMC genes in 6 tissues and 6 fruit development stages of strawberry were carried out by qRT-PCR.  Result  The 21 FaMC proteins identified in this study could be divided into Type-I, Type-I*, and Type-II based on the differences on their sequences. The phylogenetic trees and conserved domains of the 3 different types showed the differentiations between them. Type-I FaMC protein consisted of N-terminal domain (NTD) with one zinc finger motif, Type-I* consisted of NTD but no zinc finger, and Type-II had not NTD. The FaMC genes were distinctively differently expressed in different tissues and developmental stages. Type-I expressed in the leaves, Type-I* in the roots, and Type-II in the flowers. And, during the berry development stage, most Type-I* exhibited a high-low-high U-pattern of changes, whereas, part of Type-I and Type-II had a reversed U-pattern.  Conclusion  FaMC genes were identified to be closely related to the synthesis of metacaspases that played important roles in the development and growth of strawberry plants.
Effect of CO2 Enrichment on Photosynthesis of Melon Leaves in a Life Cycle
LI Tian, ZHANG Zhipeng, HOU Leiping, SUN Sheng, XING Guoming
2020, 35(11): 1198-1206.   doi: 10.19303/j.issn.1008-0384.2020.11.004
[Abstract](67) [FullText HTML](41) [PDF 1158KB](0)
Abstract:
  Objective  Changes on photosynthesis of leaves of muskmelon vines grown by hanging in a greenhouse enriched with CO2 were studied to understand the effect on the plants in a life cycle and application for optimum trellis cultivation.   Method  Four CO2 enrichment treatments were implemented in the experimentation. They included CK at 400±12 mol·mol−1, T1 at 800±24 mol·mol−1, T2 at 1 200±36 mol·mol−1, and T3 at 1 600±48 mol·mol−1. Changes on the chlorophyll content, photosynthetic indicators, and chlorophyll fluorescence indices of the leaves on the hanging melon vines during the entire growth period were determined in 5, 15, 25, 35, and 45d after the treatments.  Result  Compared with CK, T2 and T3 increased the net photosynthetic rate by 51.3% and 45.7% and WUE by 58.0% and 114.0%, respectively; while reduced the regulatory energy dissipation quantum yield Y (NPQ) by 7.1% and 11.0%, and the non-photochemical quenching coefficient qN by 5.8% and 6.2%, respectively. For T3, the increase on photochemical quenching coefficient qP was 3.9%. As the leaves aged (i.e., 45-d-old at end of treatment), the net photosynthetic rate under CK was only 19.2% at peak, whereas, that under treatments remained at a higher level. T3 had its net photosynthetic rate increased by 469.2%, chlorophyll A content significantly increased by 20.6%, Y(II) increased by 16.3%, qP increased by 25.1%, and Y(NO) decreased by 13.3% over CK. T1 and T3 showed their stomatal conductance reduced by 35.1% and 31.9%, transpiration rate by 52.8% and 37.7%, and Y(NPQ) by 9.1% and 6.6%, respectively. And, on qN, T1, T2, and T3 decreased by 8.3%, 14.6%, and 1.1%, respectively.  Conclusion  The photosynthetic capacity of 25-d-old muskmelon leaves reached the maximum with the optimal CO2 enrichment at 1 200 mol·mol−1 under T2. The enrichment in the atmosphere also increased WUE and the photochemical quenching with a reduction on the non-photochemical quenching of the plants. The combined effect would improve the efficiency of leaf photosynthesis during plant senescence when the biochemical process depends on the increased chlorophyll content and qP to function well.
Effect of Plant Grafting on Flavoring Volatiles in Large-fruit Tomatoes
LIU Ziji, LIU Weixia, NIU Yu, YANG Yan
2020, 35(11): 1207-1214.   doi: 10.19303/j.issn.1008-0384.2020.11.005
[Abstract](0) [FullText HTML](0) [PDF 603KB](0)
Abstract:
  Objective   Effect of grafting on flavoring volatiles in the large-fruit tomatoes was determined to study the flavor regulation of the plant.   Method   The composition and contents of volatiles in the fruits of the non-grafted (S) and grafted (J) large-fruit varieties, Boshou No. 1, Huiteng, and Huiteng No. 2, were analyzed for comparison using the headspace solid phase micro-extraction with gas chromatography-mass spectrometry.   Result  There were 59 including 7 unique compounds in the fruits from the Boshou No.1S plants, and 62 with 10 unique compounds in the fruits from the Boshou No.1J plants. On Huiteng S, 59 including 8 unique volatiles, and on Huiteng J, 64 including 13 unique volatiles were identified. For Huiteng No. 2S, 58 including 7 unique compounds were found, while in Huiteng No.2J, 60 including 9 unique compounds were identified.   Conclusion  The contents of volatile flavoring compounds, 6-methyl-5-hepten-2-one, β-ionone, and E-2-hexenal, were significantly higher in the fruits from the grafted (J) than the non-grafted (S) plants of 3 large-fruit tomato varieties.
Optimization of Media for Seed Germination and Propagation of Arundina graminifolia
XIA Chunying, XIE Xiaomin, LIU Jiangfeng, ZHENG Chengle
2020, 35(11): 1215-1221.   doi: 10.19303/j.issn.1008-0384.2020.11.006
[Abstract](75) [FullText HTML](109) [PDF 662KB](3)
Abstract:
  Objective   Formulation of culture media was optimized to facilitate the seed germination, tissue propagation, and various developmental stages of Arundina graminifolia.   Method   An orthogonal experiment was conducted to optimize the media for aseptic germination of the seeds and development of the seedlings of A. graminifolia using 1/2 MS as the base. Application of growth regulators and additives in the solid medium formulation was evaluated in accordance to the seed germination, seedling growth, root development and transplanting efficiency.   Result   The optimum media for various developmental stages of A. graminifolia were determined to include (1) 1/2 MS + 7 g agar·L−1 + 30.0 g sucrose·L−1 + 1 g activated carbon·L−1 + 100 mL coconut milk·L−1 + 1.0 mg NAA·L−1 + 2.0 mg 6-BA·L−1 + 0.5g Huabao No. 1·L−1 to reach a seed germination rate of 83.79% . (2) 1/2 MS + 7 g agar·L−1 + 3 0g sucrose·L−1 + 1 g activated carbon·L−1 + 1 mg IBA·L−1 + 3 mg 6-BA·L−1 + 0.5 mg NAA·L−1 + 100 g mashed potato·L−1 to achieve an average bud proliferation rate of 1.36. (3) 1/2 MS + 7 g agar·L−1 + 30 g sucrose·L−1 + 1 g activated carbon·L−1 + 0.2 mg IBA·L−1 + 2 mg 6-BA·L−1 + 0.2-0.5 mg NAA·L−1 + 100 g banana puree·L−1 for an optimal root development. (4) Perlite:peat soil mixture at 1 1 ratio to yield a seedling survival rate of 98% with desirable transplanting and acclimatization results.   Conclusion   Coconut milk in culture medium was found to significantly benefit the seed germination. The individually formulated media for various growth stages of A. graminifolia seedlings would help ensure the success in preserving the wild germplasms.
Effect of Nighttime Exposure to Light from Color Lamps on Cold Tolerance of Tomato Seedlings
LIU Fuchang, YANG Yukai, LIN Biying
2020, 35(11): 1222-1227.   doi: 10.19303/j.issn.1008-0384.2020.11.007
[Abstract](105) [FullText HTML](72) [PDF 913KB](1)
Abstract:
  Objective   Effect of exposure to light of varied spectra at night on the cold tolerance of tomato (Lycopersicon esculentum) seedlings was investigated.  Method  Tomato seedlings grown under supplemental lighting at night by using white (W), red (R) or blue (B) light or combined 2R/8B, 5R/5B or 8R/2B were taken as treatments, and seedlings grown under natural conditions were taken as control. After 20d of cultivation, the plants were subjected to a low-temperature stress of 10/5 ℃ (day/night temperatures) to determine the tolerance of the seedlings toward mild hypothermia.  Result   The ranges of increased electrolyte permeability and MDA content in the leaves of the treatment seedlings under low-temp stress were significantly lower than those of control. After being exposed at night to the combination of 2R/8B, 5R/5B or 8R/2B, the seedlings could maintain high activities of SOD, CAT, and POD under the low-temp stress. These activities peaked when the 8R/2B was applied to reach significantly 399.49% on SOD, 129.01% on CAT, and 135.74% on POD higher than those of CK, 6d after the seedlings were moved to the low-temp environment.   Conclusion  Exposing tomato seedlings to light of different spectra at night improved the ability of the plants to tolerate mild hypothermia. The effect was maximized when the seedlings were grown for 20d under the nighttime lighting with 8 red and 2 blue lamps.
Plant Protection
Transformation of Green Fluorescent Protein of Fusarium oxysporum Isolated from Diseased Bitter Gourd
CHEN Yanping, LIU Xin, XIAO Rongfeng, ZHU Yujing, Lin Yongsheng, LIU Bo
2020, 35(11): 1228-1233.   doi: 10.19303/j.issn.1008-0384.2020.11.008
[Abstract](91) [FullText HTML](73) [PDF 759KB](0)
Abstract:
  Objective  As a genetic marker, the green fluorescent protein gene (gfp) of Fusarium oxysporum f. sp. momodicae (Fom), one of the most serious fungal pathogens that caused the fusarium wilt on bitter gourd, was identified and transformed into bacterium to facilitate the study on the infection process and control of the disease.  Method  A highly pathogenic wild-type strain, FJAT-3018, isolated from diseased bitter gourds was transformed with the gene encoding gfp using an Agrobacterium-mediated method. The genetically stable transformants, FJAT-31290 and FJAT-31284, were verified by their colony morphology, growth rate, and pathogenicity.   Result  The efficiency of transforming FJAT-3018 was approximately 14.5 transformants per 106 spores. After 10 generations of subculture, the transformants did not significantly differ from the wild-type strain with respect to the colony morphology, growth rate, and pathogenicity. Strong constitutive expression of gfp could be seen in the fungal hyphae and conidia of the transformants. In addition, under a confocal laser scanning microscope, the transformant-infected bitter gourd plants showed growth of the fungal hyphae inside the roots and stem xylem.   Conclusion  The gfp gene had been successfully transformed into FJAT-31290 and FJAT-31284 with genetic stability and without apparent pathogenicity deviation from the highly pathogenic wild-type strain FJAT-3018.
Diversity of Bacillus and Fusarium Species in Rhizosphere Soil under Continuous Achyranthes bidentata Monoculture
CHEN Ting, WANG Juanying, WU Linkun, CHEN Jun, WU Hongmiao, LIN Sheng, LIN Wenxiong
2020, 35(11): 1234-1243.   doi: 10.19303/j.issn.1008-0384.2020.11.009
[Abstract](225) [FullText HTML](109) [PDF 1110KB](3)
Abstract:
  Objective   Changes on the key microbial communities in the continuously cropped rhizosphere soil (CCRS) of Achyranthes bidentata Blume were studied to analyze the allelopathic effect for operational improvement on cultivation of the medicinal plants.   Method  The community structures of Bacillus spp. and Fusarium spp. in the rhizosphere soil of A. bidentata under 1, 10, and 15 years of consecutive monoculture were determined using denaturing gradient gel electrophoresis (DGGE). Soil sample from a virgin land was used as control (CK).   Result   Abundant Bacillus spp. dominated by B. stubtilis and B. cereus were isolated from the CCRS specimens. The CCRS under 10-year (10Y) and 15-year (15Y) of continuous cropping had a greater relative content of B. subtilis and B. halodurans than their 1-year (1Y) counterpart. With respect to Fusarium spp., the number of bands and species were relatively few, the community structure tended to be similar between CK and 15Y as well as between 1Y and 10Y, and F. solani and F. oxysporum being the dominant species. The qPCR results also indicated that continuous monoculture of A. bidentata increased the abundance of the beneficial bacteria but kept that of the harmful Fusarium spp. largely unchanged.   Conclusion   Continuous cropping A. bidentata enhanced the diversity and abundance of the beneficial Bacillus spp. without encouraging the expansion of the pathogenic Fusarium community in the rhizosphere. It appeared that the rhizosphere allelopathy under the cultivation practice had created a healthy ecosystem that promoted the growth of A. bidentata.
Animal Science
Proteomes of Sheep Alveolar and Mouse Macrophages Infected by Mycoplasma ovipneumoniae
ZHANG Ying, ZHANG Kai, MA Jincheng, LI Xiaojie, MA Chunji, GAO Liyang
2020, 35(11): 1244-1251.   doi: 10.19303/j.issn.1008-0384.2020.11.010
[Abstract](95) [FullText HTML](53) [PDF 1169KB](2)
Abstract:
  Objective  Feasibility of using mouse macrophage cell line Raw 264.7 for studies on pathogenesis of Mycoplasma ovipneumoniae(Mo) on sheep was investigated by comparing the proteome changes taken place in both macrophages after an induced Mo infection.   Method   The primary alveolar macrophages from sheep lung tissue were isolated and used, along with Raw 264.7, for the study. Both macrophages were infected with a standard strain of Mo (MOI=10) for 24h prior to proteomic and RT-PCR analyses to compare the expressions on the target genes in them.   Result   The image of specific surface antigen CD14 shown on the cells obtained from the lung tissue confirmed the identity of the isolated sheep primary alveolar macrophages. After infected by Mo, both sheep and mouse macrophages showed a similar significant change pattern on the expressions of FADD, IL-1β, NOS2, and THBS1. These proteins are known to associated with the Toll-like receptor signaling pathway, MAPK signaling pathway, and autophagy in biological processes.  Conclusion  It appeared that Raw 264.7 could be satisfactorily used to substitute the sheep primary alveolar macrophage in a simulated system for studies on Mo pathogenesis to considerably simplify the process and save cost in conducting the experiments on sheep.
Factors Affecting Reproductive Performance of Saba Sows
LI Mingli, ZHAO Bao, LI Zhijuan, WANG Xiaoyi, YAN Dawei, REN Wenhui
2020, 35(11): 1252-1257.   doi: 10.19303/j.issn.1008-0384.2020.11.011
[Abstract](111) [FullText HTML](81) [PDF 569KB](5)
Abstract:
  Objective   Reproductive performance of Saba sows affected by the animal breed, parity, mating, and farrowing were investigated to explore means for improvements.  Method  From the 2013-2017 records maintained at Chuxiong Prefecture Pig Breeding Farm in Yunnan province on Saba sows that delivered 182 litters, Duroc×Saba sows (DS) that delivered 67 litters, and Landrace×Saba sows (LS) that delivered 67 litters, the reproductive performances of these sows were compared. The number of parities, insemination methods, and farrowing seasons of the litters by the sows were analyzed using the least square model.  Result  Compared with the Saba sows, DS and LS had 2.56 and 1.98, respectively, more total number born (TNB) (P<0.01) and 2.36 and 1.99, respectively, higher number born alive (NBA) (P<0.01) per litter. In general, the 4th to the 6th parities performed better than the others (P<0.05 or P<0.01), and the highest TNB and NBA occurred on the 6th while the lowest on the 1st and 9th parities. No significant differences were found on the performance whether the sows were impregnated by natural service (NS) or artificial insemination (AI) (P>0.05). However, farrowing in spring and summer rendered more desirable results than in autumn and winter (P<0.05 or P<0.01).   Conclusion  The reproductive performance of Saba sows could be improved by cross-breeding and/or farrowing in spring and summer in a year.
Utilization of Fermented Mulberry Leaves to Enhance Ruminant Digestion of Hu Sheep
LUO Yang, HE Fang, HUAN Cheng, LI Haobang, SUN Ao, LI Jianbo, LI Sheng, YI Kangle
2020, 35(11): 1258-1264.   doi: 10.19303/j.issn.1008-0384.2020.11.012
[Abstract](114) [FullText HTML](98) [PDF 634KB](0)
Abstract:
  Objective   An in vitro gas production and nutrient degradation test was applied to determine the possibility of using fermented mulberry leaves in feed for Hu sheep.   Method   Three healthy adult Hu sheep were fed each on alfalfa, Leymus chinensis or fermented mulberry leaves for comparison in this study. Ruminant liquids from the sheep were extracted and designated as ARL (from the sheep fed on alfalfa), LRL (from the sheep fed on L. chinensis), and MRL (from the sheep fed on the fermented mulberry leaves) for the 48 h in vitro simulated fermentation culture test. Production of selected gases and degradation of dry matters and fibers from the test were measured in 48-hour intervals during the fermentation process.   Result   After 48h of the in vitro fermentation, the total gas produced by MRL was 187.25 mL·g−1, which was significantly higher than LRL (P<0.01) but not with respect to ARL (P>0.05); the 46.78 mmol·L−1 total volatile fatty acids (TVFA) significantly higher than either LRL or ARL (P<0.01); the acetate, propionate, butyrate, valerate, isobutyrate and isovalerate, with the exception of propionate, concentrations of 29.76, 10.18, 4.12, 1.54, 0.48, and 0.70 mmol·L−1, respectively, significantly higher than LRL (P<0.01); butyrate and valerate significantly higher than ARL (P<0.01); and, the ratio of
acetate to propionate is 2.92 significantly higher than LRL or ARL (P<0.01). There were extremely significant differences on NH3-N content among the fermentation broths of MRL, ARL, and LRL at 1.03, 1.75, and 0.71 mmol·mL−1, respectively (P<0.01). The in vitro dry matter digestibility (IVDMD) by MRL of 46.76% was extremely significantly lower than the others (P<0.01), but the neutral detergent fiber digestibility (IVNDFD) of 70.25% significantly higher (P<0.01).
  Conclusion   The in vitro fermentation test with the ruminant liquids employed in this study clearly demonstrated the enhancing effect of MRL in the gas production and nutrient degradation as compared to ARL or LRL. It appeared that the fermented mulberry leaves could be applied in feed to promote the ruminant digestion of Hu sheep.
Resources and Environmental Science
Intein-mediated Expression in E. coli and Amidation of Antimicrobial peptide MME
YE Ruobai, WU Zhenhong, MIAO Xiaoqing
2020, 35(11): 1265-1270.   doi: 10.19303/j.issn.1008-0384.2020.11.013
[Abstract](0) [FullText HTML](0) [PDF 917KB](0)
Abstract:
  Objective   Efficient method to amidate the carbon-terminal of the antimicrobial peptide MME through self-splitting intein in recombination fusion proteins expressed in E. coli with DTT in the presences of MESNa and NH4HCO3 was explored.   Method   Recombinant plasmid of MME was constructed to induce the intein-mediated expression in E. coli to successively obtain fusion proteins composed of histidine, sumo label, target polypeptide, and intein. The proteins were subsequently purified in a process using a nickel column and dialysis. With MESNa and NH4HCO3, an intein self-split procedure was completed by DTT to amidate the carbon-terminal on MME, then cut and purify the intestinal kinase. Both MME and the amidation were verified by the standard mass spectrometry and two-stage, tandem mass spectrometry.   Result   The resultant gene fragment of the fusion protein was determined by PCR to be 837bp long as expected. In comparison to the theoretical value of 3 057.64, the relative molecular weight of MME obtained by the standard mass spectrometry was 3 057.7. The molecular weight of the amidated carbon-terminal fragments of MME was measured by the two-stage, tandem mass spectrometry to be 1,214.728, which was close to the known theoretical value of 1 214.739 with a matching rate of 45. It appeared that the carbon-terminal of MME had been effectively amidated, and the protein obtained was soluble.   Conclusion   The recombinant fusion proteins were successfully expressed in E. coli to enable the intein self-splitting in the presences of MESNa and NH4HCO3 and amidate the carbon terminal of MME. Therefore, the current simple, one-step preparation could achieve accurate, duplicatable, and meaningful results. By combining the standard mass spectrometry and two-stage, tandem mass spectrometry, the amidated carbon-terminal polypeptides could satisfactorily be detected.
Ecotoxicity of Nonylphenol on Eisenia foetida and Detoxification by Tea Polyphenols
CHEN Jianhua, ZHANG Biyao, LI Danpei, CHEN Shihong, XIE Yanying, WANG Guohong
2020, 35(11): 1271-1279.   doi: 10.19303/j.issn.1008-0384.2020.11.014
[Abstract](122) [FullText HTML](88) [PDF 1054KB](1)
Abstract:
  Objective  Ecotoxicity imposed by nonylphenol (NP) on Eisenia foetida and potential detoxification effect of tea polyphenols were studied.  Method   Adding different concentrations of tea polyphenols into NP base solution, the relief of tea polyphenols (TP) on the life state, lethal toxicity, avoidance behavior and growth inhibition induced by NP were observed in E. foetida, and the toxic mechanism of NP was studied by HE staining.  Result  At a concentration in soil ranging between 100-200 mg·kg−1, NP exerted varied degrees of toxicity to the earthworms with lethality directly correlated with the concentration as well as exposure time. The growth of earthworms was retarded by 34% at 200 mg NP·kg−1, and a high mortality rate observed when the concentration reached 400 mg NP·kg−1. The stained tissues of the earthworms under NP stress showed decreased vacuolated gland cells, thinned circular muscles, bared cuticles, and thickened epidermis. Toward the soil with NP in the range of 12.5-200 mg·kg−1, the earthworms displayed a movement avoidance. Upon the addition of tea polyphenols at the concentrations of 70-700 μg·kg−1 in the NP solution, the lethality and growth retardation of the earthworms declined significantly. A tea polyphenols addition of 0.7-7 μg·kg−1 in the NP solution significantly lessened the avoiding response of the earthworms. With 0.7-700 μg·kg−1 of added tea polyphenols, the thickness of the muscular layer of the earthworm body wall increased and the stratum corneum became smooth again.  Conclusion   NP at a concentration in the range from 100 mg·kg−1 to 400 mg·kg−1 in soil could be toxic or lethal to E. foetida. Whereas, the presence of tea polyphenols at a level between 0.7 μg·kg−1 and 700 μg·kg−1 in the NP solution that was blended in the soil could significantly mitigate the adverse effect induced by NP.
Effects of Interactions between Roots of Intercropped Maize and Soybean on Plant Photosynthesis, Crop Yield, and Soil Physiochemical Properties
SHEN Lihua, LI Na, RUAN Miaohong, LIN Wenxiong
2020, 35(11): 1280-1288.   doi: 10.19303/j.issn.1008-0384.2020.11.015
[Abstract](77) [FullText HTML](45) [PDF 697KB](0)
Abstract:
  Objective   Effects of underground interactions between roots of maize and soybean plants under intercropping on plant photosynthesis, crop yield, and soil physiochemical properties were investigated to decipher the associated mechanisms.   Method  A maize/soybean intercropping experimentation was conducted in the field. Underground between the maize and soybean plants, a plastic sheet to completely separate and deprive interactions between the root systems (Q), a nylon mesh to partially block the underground interactions (W) or with no artificial barrier to allow total root-interactions (N) was implemented. SPAD, photosynthetic characteristics, and yield of the plants grown under the varied degrees of partition on the root systems were measured.  Result   Compared to monoculture, the intercropped maize and soybean under N showed increased chlorophyll contents in the functional leaves by 10.36% and 9.65%, respectively. The net photosynthesis, stomatal conductance, intercellular CO2 concentration, and transpiration rate were also significantly enhanced by the intercropping under varied roots partitions as they ranked N>W>Q>monoculture. The crop yields of the plants were improved by the intercropping under W or N. The land equivalent ratio (LER) of the N treatment was 1.39, while that of W 1.13. Under Q or W, the contents of total and available N, P, and K in rhizosphere soil were higher than those in the monoculture lot. The activities of catalase, acid phosphatase, urease, sucrase, and peroxidase increased in the rhizosphere soil under W or N. A significant correlation was observed between part of the enzyme activity and nutrient content in the soil.   Conclusion  The interactions between the root systems of the intercropped maize and soybean plants might activate the nutrient pool and enzyme activity in the rhizosphere soil. The leaf chlorophyll of the intercropped plants could also be increased by the underground interactions benefitting the plant photosynthesis with improved crop yield.
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