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    25 April 2020, Volume 41 Issue 4 Previous Issue    Next Issue
    Crop Culture and Nutrition, Genetic Breeding
    Nutrient Content of Coffee Peel with Different Composting Treatments and Its Effects on Coffee Plant Growth
    ZHAO Qingyun,PU Haojie,WANG Qiujing,DONG Yunping,LIN Xingjun,SUN Yan,LONG Yuzhou
    2020, 41 (4):  633-639.  doi: 10.3969/j.issn.1000-2561.2020.04.001
    Abstract ( 747 )   HTML ( 18 )   PDF (1413KB) ( 318 )  

    Coffee peel can be used as a potential organic fertilizer source for agricultural production, but the application of the coffee peel not rotten can inhibit the growth of coffee seedlings. Coffee peel and shattered coffee peel rotten with nothing or cellulase and pectinase were carried out to investigate the effects of different treatments on the coffee peel nutrient content and plant growth, and to provide basis for coffee peel fertilizer utilization. The results of laboratory tests showed that there was no significant difference on the nutrient content of coffee peel among the treatments, whereas the organic matter in the shattered coffee peel treated with cellulase and pectinase decreased by 7.90% compared with the coffee peel treated with no enzymes. Greenhouse pot experiments showed that the application of well-rotten coffee peel could significantly promote the growth of coffee seedlings, increase the leaf photosynthetic rate, chlorophyll content and nitrogen balance index. The application of shattered coffee peel treated with cellulase and pectinase increased the biomass production of coffee plant, leaf photosynthetic rate, chlorophyll content and nitrogen balance index over 50% compared with the control. Conclusively, shattered coffee peel treated with cellulase and pectinase was suggested to be used to promote the growth of coffee plants.

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    Comparison of Yield Formation and Quality of ‘Zhongjiao No. 9 ’ under Different Transplanting Dates
    TIAN Qinglan,LIU Jieyun,WU Yanyan,HUANG Weihua,HUANG Yongcai,MOU Haifei,WU Daidong,ZHANG Yingjun,HUANG Pingming
    2020, 41 (4):  640-648.  doi: 10.3969/j.issn.1000-2561.2020.04.002
    Abstract ( 1045 )   HTML ( 18 )   PDF (1288KB) ( 283 )  

    ‘Zhongjiao No. 9’, a new banana variety with high resistance to ‘fusarium wilt No. 4’, was introduced and transplanted in different dates from January to October in Nanning, Guangxi, China, and its growth characteristics, yield and quality were observed in order to provide references for the selection of disease-resistant varieties in Guangxi. ‘Guijiao No. 1’ was used as the control, a traditional variety in Guangxi, and the plant growth characteristics, yield and fruit quality of ‘Zhongjiao No. 9’ and ‘Guijiao No. 1’ under different transplanting dates were determined. The results showed that: ‘Zhongjiao No. 9’ had a greater yield advantage than ‘Guijiao No. 1’. The average yield of ‘Zhongjiao No. 9’ was 16.2% higher than that of ‘Guijiao No. 1’. ‘Zhongjiao No. 9’ had higher yield under transplanting date of Feb. 15stand Oct. 1st. The growth period of ‘Zhongjiao No. 9’ under different transplanting dates was 140~181 d longer than that of ‘Guijiao No. 1’, and its growth period under summer and autumn planting was 161 d longer than that under winter and spring planting on average, mainly due to bud squaring delayed. Besides, compared with ‘Guijiao No. 1’, ‘Zhongjiao No. 9’ had longer growth period, later bud squaring, higher leaf photosynthetic potential and leaf area index before budding, which were the basis of its biomass and yield formation. In terms of quality, the content of total sugar and titratable acid of ‘Zhongjiao No. 9’ under transplanting dates of Jan. 1st was 29.0% and 63.0% higher than that of ‘Guijiao No. 1’ after fruit ripening, respectively. The fat content of ‘Zhongjiao No. 9’ was significantly lower than that of ‘Guijiao No. 1’. Therefore, the fruit of ‘Zhongjiao No. 9’ after ripening showed high sugar, acid and low fat. In conclusion, ‘Zhongjiao No. 9’ had a certain yield advantage in Guangxi, but its growth period was longer and the bud was later than ‘Guijiao No. 1’. Transplanting on Feb. 15st in winter and spring could achieve high yield and shorten the growth period for ‘Zhongjiao No. 9’, but it is still necessary to further observe its comprehensive performance in the disease area.

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    Partial Substitution of Chemical Fertilizer by Organic Fertilizer Improving Nutrition and Fruit Quality of ‘Sanhong-miyou’
    LI Shuixiang,SHE Wenqin,WU Shitao,CHEN Jingying,MA Wen,SUN Yuchen,WANG Jie
    2020, 41 (4):  649-654.  doi: 10.3969/j.issn.1000-2561.2020.04.003
    Abstract ( 643 )   HTML ( 10 )   PDF (1178KB) ( 362 )  

    A field experiment was carried out to study the effects of organic fertilizers partly replacing of chemical fertilizers on the nutrition and fruit quality of ‘Sanhong-miyou’. Two fertilization treatments were selected: 20% of chemical fertilizer replaced by organic fertilizer (T) and only chemical fertilizer (CK). The results showed that T could significantly increase the magnesium and zinc contents in the leaves of ‘Sanhong-miyou’ and reduce the nitrogen content of the leaves to the appropriate level, increase the content of potassium, calcium, zinc and boron in fruits of ‘Sanhong-miyou’, reduce the phosphorus content during fruit development. In addition, it also significantly increased the soluble solid, total sugar and sugar acid ratio, reduced titratable acid of the fruits, and the Vc content, edible rate and single fruit quality of it did not show significant difference compared with CK.

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    Transplantation Effects of Different Nursery Substrates on the Growth of Tissue Culture Seedlings of Mytilaria laosensis
    LIU Zhenxiang,LI Rongsheng,ZOU Wentao,QIU Zhenfei,YU Niu,YANG Jinchang
    2020, 41 (4):  655-660.  doi: 10.3969/j.issn.1000-2561.2020.04.004
    Abstract ( 704 )   HTML ( 17 )   PDF (1227KB) ( 391 )  

    The effect of different nursery substrates on the transplant survival rate, growth indicator and seedling quality index of M. laosensis tissue culture seedlings was investigated by setting five different nursery substrates according to the volume ratio of two nursery substrates (yellow soil and peat soil). T1 (100% yellow soil) and T3 (60% yellow soil+40% peat soil) were more conducive to the survival of M. laosensis seedlings after transplantation, and the survival rate of the seedlings transplanted for 1 month was the highest (83.33% for T1, and 81.11% for T3). Plant height, leaf number and ground diameter of M. laosensis seedlings increased first and then decreased with the increase of the proportion of peat soil, and reached the maximum at T3. The three indicators of the seedlings transplanted after 6 months for T3 were 30.93 cm, 8.1 and 5.04 mm, respectively, which were significantly higher than those of the other four treatments (P<0.01). The aboveground and underground dry weight of M. laosensis seedlings transplanted after 6 months were T3 > T4 > T5 > T2 > T1, and the aboveground and underground dry weight for T3 were 8.51 and 5.56 times of those of T1. Furthermore, the seedling quality index of T3 reached 1.13, which was significantly higher than that of T5, T2 and T1 (P<0.05). Therefore, the mixture nursery substrates with 60% yellow soil and 40% peat soil were the ideal nursery substrates for tissue culture seedlings of M. laosensis.

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    Relationship Between SRAP Marker Based on Genetic Distance, Combining Ability and Heterosis in Pepper
    WU Lidong
    2020, 41 (4):  661-668.  doi: 10.3969/j.issn.1000-2561.2020.04.005
    Abstract ( 607 )   HTML ( 7 )   PDF (1015KB) ( 228 )  

    The SRAP was used to analyze the correlation between the geneic distance with combining ability and heterosis of various agronomic traits in pepper. The materials consisted of 11 parent peppers and 30 F1 hybrids based on half diallel methods. The results showed that there was no significant correlation between the genetic distance and general combining ability (GCA), special combining ability (SCA) and heterosis of pepper parents. There was no significant correlation between SCA and heterosis of all traits, but only significant positive correlation between heterosis of fruit length and GCA. It is difficult to predict the combining ability and heterosis of the offsprings by the genetic distance, GCA and SCA could not be used to predict heterosis completely. Therefore, the relationship between the genetic distance, combining ability and heterosis needs to be further studied.

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    Establishment of Coix lacryma-jobi L. Core Germplasm Collection Based on Phenotypic Characters
    LI Xiushi,FU Yuhua,ZHOU Xiang,LI Qing,LIU Fanzhi,YANG Chenglong,ZHOU Mingqiang
    2020, 41 (4):  669-675.  doi: 10.3969/j.issn.1000-2561.2020.04.006
    Abstract ( 566 )   HTML ( 7 )   PDF (1044KB) ( 278 )  

    A total of 248 accessions of Coix lacryma-jobi L. were grouped by geographical source and least distance stepwise sampling based on 14 phenotypic data. Eight candidate core collections were obtained according to a certain sampling proportion. The candidate core collections were then evaluated by the variable rate of coefficient of variation, mean difference percentage, coincidence rate of range, variance difference percentage, ratio of phenotypic retained, variance of phenotypic frequency and Shannon-Wiener diversity index. The primary core collections of C. lacryma-jobi L. contained 67 accessions with 27.02% of the original germplasm resources retained (25% sampling ratio within the group). The variable rate of coefficient of variation, mean difference percentage, coincidence rate of range, variance difference percentage, ratio of phenotypic retained, variance of phenotypic frequency and Shannon-Wiener diversity index was 110.05%, 0, 93.03%, 10%, 96%, 0.655, 0.881, respectively. Results of t-test showed that no significant difference was found in the mean of 14 phenotypic data between the primary core collections and original collections, with only three phenotypic traits lost some trait levels, 11 phenotypic traits retained all trait levels of the original collection (RPR=1.0), and the proportion of retained phenotypic data levels was 96.49%. The variation coefficient (CV) and Shannon-Wiener diversity index (I) of most traits were slightly higher than those of the original collection. It indicated that the core collection was of more representative and richer genetic diversity. The results laid an important foundation for the collection, protection and innovation of C. lacryma-jobi L. germplasm resources.

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    Phytophysiology and Biochemistry
    Germination Characteristics of 85 Introduced Stylosanthes Seed under PEG Stress
    ZHANG Yu,YAN Linling,YU Daogeng,WANG Wenqiang,LIU Guodao
    2020, 41 (4):  676-684.  doi: 10.3969/j.issn.1000-2561.2020.04.007
    Abstract ( 596 )   HTML ( 9 )   PDF (1222KB) ( 267 )  

    In order to investigate the drought resistance of different Stylosanthes, an evaluation system for the drought resistance of Stylosanthes resources was constructed. Using 85 imported Stylosanthes as the test material, the seed relative germination rate, relative germination energy, relative radiele length, relative plumule length, germination drought resistance index were measured under distilled water (CK), -0.2 and -0.6 MPa PEG-6000 stress. The drought resistance ability of the germplasms was evaluated comprehensively by the subordinate function method by analyzing the effects of different drought stress on the seed germination of Stylosanthes. Low concentration of PEG had a certain promotion on the germination of seeds. High concentration of PEG reduced the relative germination potential and relative germination rate of Stylosanthes seeds, hindered the growth of embryo and radicle, and showed great differences among species. Among them, drought resistant materials were S. capitata CIAT 2250, S. macrocephala M.B. CIAT 2113 and S. macrocephala M.B. CIAT 1942, S. montevidensis Vogel CIAT 10182, and non-drought resistance materials were S. mexicana Taub. CIAT 1590, S. calcicola Small CIAT 1616 and S. calcicola Small CIAT 1624. The result of the study would provide a theoretical basis for the drought resistance evaluation of introduced resources, breeding of drought resistant varieties and drought resistance mechanism research of Stylosanthes.

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    Analysis of Physio-biochemical Characteristics of T2 α-tubulin SoTUA Transgenic Sugarcane
    CHEN Jiaoyun,KHAN Qaisar,WEI Jianglu,TANG Lihua,DONG Dengfeng,LI Yangrui
    2020, 41 (4):  685-693.  doi: 10.3969/j.issn.1000-2561.2020.04.008
    Abstract ( 628 )   HTML ( 6 )   PDF (1381KB) ( 302 )  

    Six SoTUA transgenic lines (T1, T2, T3, T4, T5, T6) were selected from the positive SoTUA transgenic sugarcane lines of T2 generation after PCR amplification and sequencing. The transgenic lines and control (wild type, WT) were treated with low temperature stress (4 ℃), and the level of malondialdehyde (MDA) and osmotic adjustment substances (soluble sugar, soluble protein) and activities of peroxidase (POD) and superoxide dismutase SOD were determined, and RT-qPCR analysis was done for SoTUA gene expression in sugarcane leaves at 0, 5 and 10 days of the treatment. The relative expression of α-tubulin transcription factor in the SoTUA transgenic sugarcane and WT was up-regulated, getting higher with the cold stress time under low temperature treatment at 4 ℃. The relative expression level was always higher in the SoTUA transgenic lines than that in WT at 5 and 10 days of the treatment. At the 10th day, the relative expression level of the transgenic line T1 was significantly higher than that of other transgenic strains (P<0.05). The content of MDA increased gradually under cold stress, but it was found significantly lower in the transgenic sugarcane than that in WT by 41.77% (P<0.05). The trend of POD activity varied among different lines, but it was higher in the SoTUA transgenic lines than in WT (P<0.05). The SOD activity of transgenic lines T1, T2, T4 and T6 increased first and then decreased while SOD activity of WT and transgenic lines T3 and T5 increased gradually. The soluble sugar content increased gradually and the trend of soluble protein varied in different strains. The expression of SoTUA gene was positively correlated with soluble sugar. Using the membership function comprehensive analysis method, the ranking result for cold resistance was obtained as T3 > T2 > T1 > T5 > T6 > T4 > WT. Up-regulated expression of SoTUA gene in the transgenic sugarcane could improve cold resistance, and the genetic stability is good.

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    Silicon and Nitrogen Improving the Eco-physiological Characteristics of High Quality Rice
    PAN Taowen,CHEN Yu,CAI Kunzheng
    2020, 41 (4):  694-700.  doi: 10.3969/j.issn.1000-2561.2020.04.009
    Abstract ( 605 )   HTML ( 8 )   PDF (1273KB) ( 236 )  

    Two rice varieties ‘Guinongzhan’ and ‘Huanghuazhan’ were used as the material to study the effects of N and Si application on rice plant growth and physiological traits. Three levels of nitrogen 120, 180 and 240 kg/hm 2, indicated by N1, N2 and N3, respectively, and three levels of silicon application 0, 225 and 450 kg/hm 2, indicated by Si1, Si2 and Si3, respectively, were selected. The interaction of silicon and nitrogen could increase the panicle number, plant height and biomass of rice. The N2 and Si2 treatment significantly improved the photosynthesis of leaves, especially for ‘Guinongzhan’. The photosynthetic rate increased by 19.21% and 8.75%, respectively. Under different N application levels, medium Si level (Si2) significantly increased the nitrate reductase (NR) activity of leaves. Compared with Si treatment, Si2 treatment increased NR by 46.82% and 85.43% for ‘Guinongzhan’ and ‘Huanghuazhan’, respectively. NR activity increased with the increase of Si level at maturity stage under low N level. For sucrose invertase (SI) activity, both varieties showed the trend of different growth stage: heading stage > tillering stage > mature stage, and SI activity in ‘Huanghuazhan’ was higher than that in ‘Guinongzhan’. The results suggested that low N and high Si, medium N and medium Si treatment could significantly promote rice growth and metabolism.

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    Effects of Different Gradient Light Intensity on Healing Growing and Quality of Grafted Cucumber Seedlings
    LIAO Ziyue,LIN Biying,WANG Yue,SHEN Baoying,ZHONG Luming
    2020, 41 (4):  701-708.  doi: 10.3969/j.issn.1000-2561.2020.04.010
    Abstract ( 673 )   HTML ( 10 )   PDF (1322KB) ( 343 )  

    In order to explore the best light pattern of cucumber grafted seedling healing and seedling growth, the pumpkin cultivar ‘Zhuangshi’ was used as the rootstock and cucumber cultivar ‘Dapengdongqing’ as the scion. A stepwise incremental illumination mode was used for the seedling 0-3, 4-6, 7-10 d after top grafted. The light intensity was T1 (15-30-45) μmol/(m2·s), T2 (30-60-90) μmol/(m2·s), T3 (45-90-135) μmol/(m2·s), T4 (60-120-180) μmol/(m2·s) and T5 (75-150-225) μmol/(m2·s), CK (0-30-45) μmol/(m2·s). Compared with CK, the suitable light was better than the dark condition at 0-3 d after grafted. The light intensity gradient significantly improved the joint healing effect when treated between T2-T5 (P<0.05). It is beneficial to the healing and growth of the grafted seedlings. The growth index of above-ground and under-ground parts was the best, which was best beneficial to promoting the differentiation and early maturity of grafted seedlings. The T3 and T4 treated seedlings had better growth potential, while the T2 and T3 treated was beneficial to the increase of photosynthetic pigment content in the grafted. The ranking of the comprehensive evaluation value of membership function under each treatment was: T3 > T2 > T4 > T5 > T1 > CK. According to the comprehensive analysis, the light intensity gradient of T3 treatment in 0-3, 4-6 and 7-10 d after grafted was (45-90-135) μmol/(m2·s), which was best beneficial to the healing growth of the grafted.

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    Effects of Two Polyamines on in Vitro Flowering of Dendrobium officinale Kimura et Migo
    LI Ru,LI Zhilin,BAI Jiankun,SHANG Zhengrui,ZHAO Liying,WANG Yuying
    2020, 41 (4):  709-714.  doi: 10.3969/j.issn.1000-2561.2020.04.011
    Abstract ( 615 )   HTML ( 6 )   PDF (1775KB) ( 408 )  

    The effects of different concentrations of exogenous putrescine (Put) and spermine (Spm) on the flowering of Dendrobium officinale Kimura et Migo were studied using the rootless plantlets in vitro. The flowering rate of D. officinale could be increased by adding appropriate amount of Put and Spm in the medium. The flowering rate of D. officinale was the highest (30.47%) when the concentration of Put was 0.4 mg/L, and the flowering rate of D. officinale was 22.26% when the concentration of Spm was 0.2 mg/L. The time to have the first flower bud appeared of D. officinale was the shortest (83.33 d) when the concentration of Put was 0.2 mg/L, and the viewing period of D. officinale could be prolonged by 43.33 days. When the concentration of Put was 0.4 mg/L, the content of soluble sugar and soluble protein was the highest. The content of total N was the highest in CK. When the concentration of Spm was 0.6 mg/L, the C/N ratio of the plant was the highest. The concentration of Put 0.4 mg/L would be beneficial to the accumulation of carbon and nitrogen compounds, which could promote the flowering rate of D. officinale.

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    Biotechnology and Tissue Culture
    Cloning, Bioinformatics and Expression Analysis of Mango MiCOL6 Gene
    LU Xinxi,LUO Cong,ZHANG Xiujuan,YU Haixia,LIU Yuan,HE Xinhua
    2020, 41 (4):  715-721.  doi: 10.3969/j.issn.1000-2561.2020.04.012
    Abstract ( 707 )   HTML ( 15 )   PDF (2686KB) ( 303 )  

    CONSTANS (CO) is a core regulator of the photoperiod pathway, which integrates light quality and clock output signals to regulate the floral transformation in plants. A full-length cDNA of MiCOL6 gene was obtained from transcriptome data analysis and then verified by RT-PCR amplification in mango. Sequences analysis showed that the open reading frame was 678 bp in length, coding 226 amino, the molecular weight and isoelectric point (pI) of MiCOL6 was 26.88 kDa and 4.85, respectively; and MiCOL6 was a non-secreted hydrophilic protein. Conserved domain and phylogenetic tree analysis showed that MiCOL6 contained only one CCT domain and belonged to the group IV of the CO gene family. The secondary structure analysis showed that the protein mainly consisted of random curl and α-helix. The subcellular localization prediction showed that MiCOL6 was likely to be localized in the cytoplasm membrane. Expression pattern analysis showed that the MiCOL6 was the highest expressed in leaves during the bud differentiation period. The results of the experiment would provide a theoretical basis for the further study on the function of MiCOL6 in mango.

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    Development, Characterization and Speciality of Microsatellite Markers in AP85-441 and R570 Genomic Reference Sequences
    XU Zhijun,ZHAO Sheng,HU Xiaowen,KONG Ran,SU Junbo,LIU Yang
    2020, 41 (4):  722-729.  doi: 10.3969/j.issn.1000-2561.2020.04.013
    Abstract ( 830 )   HTML ( 17 )   PDF (2583KB) ( 298 )  

    Lacking of molecular markers is an important factor restricting the development of marker-based genetic and breeding research. In this study, 512 835 and 97 839 microsatellite sequence, which accounted for 0.32% and 0.35% of the genome, were identified based AP85-441 and R570 genomic reference sequences respectively, using the MISA software. In the genomes, the dominant repeat units were single nucleotide, dinucleotide and trinucleotide, and each repeat unit was dominated by AT-enriched elements. 472 117 and 89 748 loci of the genomes could be used to develop SSR markers. 16 691 and 13 271 homologous genes corresponding to sorghum chromosome 1-10 were identified by homology analysis on the genes of Saccharum spontaneum, sugarcane and sorghum, and 13 224 and 7624 pairs of SSR primers were developed by the gene sequence. In silico PCR analysis of these SSR markers revealed that the markers were mostly multilocus amplified in Saccharum spontaneum genome, and the effective markers in the two genomes were 79.35% and 36.13%, 79.01% and 93.36%, respectively. Part of the SSR markers showed specific amplification in the genomes. 1368 and 1420 SSR were specific single-locus amplification in AP85-441 and R570, respectively. 752 SSR were single-locus amplification in the two genomes, and the amplification sites and source genes of these SSR were distributed on all chromosomes in the genome. In this study, the SSR loci identified would be conducive to enriching the molecular markers of sugarcane, and the 3540 pairs of SSR primers developed will provide support for the determination of homologous linkage groups and group genetic origin in the construction of sugarcane genetic map.

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    Clonging and Expression Analysis of WRKY52 Gene in Dimocarpus longan
    XUE Xin,ZHOU Xinzhi,JUE Dengwei
    2020, 41 (4):  730-736.  doi: 10.3969/j.issn.1000-2561.2020.04.014
    Abstract ( 805 )   HTML ( 13 )   PDF (9812KB) ( 258 )  

    WRKY transcription factors plays an important role in plant growth, development and stress response. Based on a previous study, we found several WRKY genes, such as DlWRKY52, were participated in the process of floral induction and stress response. To further reveal the function of longan WRKY genes, DlWRKY52 was cloned using the leave cDNA of ‘Sijimi’ longan as the template. Meanwhile, the sequence characteristics, tissue expression patterns, flower and fruit development process expression patterns and subcellular localization were also studied. Bioinformatics analysis indicated that the complete open reading frame (ORF) box of DlWRKY52 was 918 bp, encoding 305 amino acid residues. The amino acid sequence alignment analysis showed that DlWRKY52 contained a typical WRKY domain and a zinc finger structure, belonging to Group Ⅱc. The result of qRT-PCR showed that DlWRKY52 was highly expressed in leave, stem and fruit organs, significantly up-regualted in the pulp 80 days post-anthesis. The transient expression of Arabidopsis protoplasts demonstrated that DlWRKY52 protein was localized to the nucleus, indicating that DlWRKY52, as a typical transcription factor, is localized to the nucleus, and might participate in the regulating of longan floral induction and early fruit development.

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    Cloning and Expression Analysis of 4-coumarate: coenzyme A Ligase Gene (Pn4CL) in Piper nigrum
    FAN Rui,HU Lisong,WU Baoduo,HAO Chaoyun
    2020, 41 (4):  737-744.  doi: 10.3969/j.issn.1000-2561.2020.04.015
    Abstract ( 596 )   HTML ( 10 )   PDF (9437KB) ( 246 )  

    The full-length cDNA encoding 4-coumarate:coenzyme A ligase (4CL), designated as Pn4CL, was isolated from black pepper using RACE. The sequence of Pn4CL was 2130 bp in length, containing a 1638 bp open reading frame, and encoding a polypeptide of 545 amino acids with a calculated molecular weight of 59.57 kDa and a PI of 5.70. The protein had four conserved domains of AMP-binding (AMP-binding enzyme), CaiC (Acyl-CoA synthetase (AMP-forming)/AMP-acid ligase), PLN02246, AFD-class I. Pn4CL had a closer relationship with Asarum heterotropoides and far from the evolution of Rosids by phylogenetic analysis. The subcellular localization indicated that the protein was on the cell membrane. The expression of Pn4CL could be regulated by exogenous MeJA and SA. When infected with Phytophthora capsici Leon, the expression of Pn4CL was significantly higher in the resistant germplasm than that in the susceptible germplasm by real-time RT-PCR.

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    Cloning and Expression Analysis of HSP70 in Oncidium hybridum
    FENG Baoyun,LI Rong,LAI Zhongxiong,LIN Yuling
    2020, 41 (4):  745-754.  doi: 10.3969/j.issn.1000-2561.2020.04.016
    Abstract ( 667 )   HTML ( 17 )   PDF (11580KB) ( 481 )  

    In order to study the molecular characteristics of the heat shock protein gene (named OnHSP70) and its expression patterns in O. hybridum ‘Honey Angel’, OnHSP70 was cloned by RT-PCR, its molecular characteristics was analyzed by bioinformatics, and its expression patterns in different tissues and under different abiotic stress treatment were carried out by qRT-PCR. Bioinformatics analysis showed that the open reading frame of the gene was 1944 bp, encoding 647 amino acids, and the translated protein was a stable protein belonging to the HSP70 superfamily. The secondary structure of the protein consisted of 41.11% α-helix, 17.77% extended chain, 7.73% β-turn and 33.38% random coil, and had two highly conserved functional domains. Clustering analysis showed the protein had closely relative to HSP70 from Dendrobium catenatum and Apostasia shenzhenica. qRT-PCR analysis showed OnHSP70 differentially expressed in four tissues and the highest expression level was found in flowers; the expression of OnHSP70 gene increased significantly after high temperature treatment, and reached the peak under 40 ℃ for 4 h treatment; OnHSP70 showed down-regulated under methyl jasmonate and salicylic acid treatment. The study would provide a theoretical basis for studying the gene function and improving the adaptability to high temperature of Oncidium hybridum.

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    Technology for Rapid Propagation in Vitro of Ancient Four-season Rhododendron Tree in Longyuan of Pingnan
    HU Jihong, CHEN Guixin, YANG Huiting, KANG Jianban, GAN Daikui
    2020, 41 (4):  755-763.  doi: 10.3969/j.issn.1000-2561.2020.04.017
    Abstract ( 584 )   HTML ( 10 )   PDF (1776KB) ( 217 )  

    Rapid propagation in vitro was explored using the apical buds of an ancient Four-season Rhododendron tree, more than 400 years, from Longyuan Village, Tangkou Town, Pingnan County as the explants to find the effects of sterilizing methods and time of explants on the development of sterile cultures, effects of kinds and ratios of plant growth regulators added in the media on the proliferation of shoots and rooting of rootless shoots. The optimum sterilization conditions were the explants treated with 75% of ethanol for 30 seconds and then dipped with 0.1% of HgCl2 for 8 minutes. The optimum sampling period for explants was about middle to late May. After four times of bottle transfer success rate of re-sterilizing reached 71% through rescuing of the contaminated explants in the development of sterile cultures by treating with 75% of ethanol for 20 seconds and then dipping with 0.1% of HgCl2 for 5 minutes. The optimum components of the medium in the primary culture was WPM medium added 3.0 mg/L of zeatin (ZT), 0.1 mg/L of NAA and 0.5 mg/L of GA3, on which the average number of induced valid buds was 3.45. In the proliferation culture, the clustered shoots were induced from apical and stem segments with axillary buds of sterile plant-lets inoculated on Anderson medium added 0.5 mg/L of TDZ and 0.1 mg/L of NAA, multiplication coefficient of stem segments with axillary buds was higher than that of apical buds, reached 13.23. In the rooting culture of rootless shoots, the optimum medium was WPM medium added 0.1 mg/L of ZT and 0.5 mg/L of NAA, on which the rooting rate reached 92.6%. The results would provide theoretical and technical supports for conservation, exploitation and utilization of the ancient Four-season Rhododendron tree.

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    Crop Pests, Diseases and Their Control, Environmental Protection
    Effects of Ultraviolet-C and Cold Treatment on Pestalotiopsis mangiferae and Physiological Indices of Postharvest Mango Fruits
    MAI Xinyun,HUANG Jiangqi,HUANG Bin,WANG Yi,LAN Xidan,LIAO Fenyan
    2020, 41 (4):  764-772.  doi: 10.3969/j.issn.1000-2561.2020.04.018
    Abstract ( 782 )   HTML ( 10 )   PDF (1416KB) ( 241 )  

    The effects of different doses of ultraviolet-C (UV-C) and storage temperature on the expansion of lesions caused by Pestalotiopsis mangiferae, total phenols, flavonoids, malondialdehyde, soluble protein, enzyme activities of peroxidase and polyphenol oxidase of postharvest mango fruits were studied. UV-C treatment at 4.9 kJ/m 2 significantly suppressed the expansion of lesions, enhanced the content of total phenols and flavonoids, reduced the content of soluble protein, increased the activity of peroxidase and polyphenol oxidase, and inhibited the production of malondialdehyde. UV-C treatment at 9.8 kJ/m 2 could damage plant cells, and weaken the promoting effects on flavonoids, soluble proteins, peroxidase and polyphenol oxidase activities. Compared with 25 ℃, low temperature treatment at 13 ℃ could suppress the expansion of lesions, maintain higher content of total phenols and flavonoids, inhibit the formation of malondialdehyde, and delay the peak time of peroxidase and polyphenol oxidase activity. UV-C treatment at 4.9 kJ/m 2 combined with storage at 13 ℃ had best a control effect on P. mangiferae of mango fruits, and maintained much better quality of postharvest mango fruits effectively.

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    Geographical Population Genetic Differentiation Analysis of Eotetranychus sexmaculatus in Rubber Tree Based on Mitochondrial COI Gene Sequences
    NIE Yue,LU Tangfei,CHEN Junyu,ZHANG Fangping,NIU Liming,FU Yueguan
    2020, 41 (4):  773-778.  doi: 10.3969/j.issn.1000-2561.2020.04.019
    Abstract ( 577 )   HTML ( 6 )   PDF (2396KB) ( 253 )  

    Eotetranychus sexmaculatus (Riley) is one of the most important pests of rubber trees, with a wide geographical distribution and heavy infestation. In order to study its genetic diversity and genetic differentiation, the mitochondrial COI gene sequence was used to analyze the genetic structure of 165 individuals from 11 different geographical populations collected from rubber trees in Hainan, Yunnan and Hanoi (Vietnam). The obtained mitochondrial COI gene fragment of the six-pointed spider mites was 658 bp in length. There were 30 variable sites, including 15 simple information points, 15 single mutation sites, and 15 haplotypes. The FST value between populations was 0.34698, the Nm value was 0.47, and the GST value was 0.19660, indicating that there was genetic differentiation in the 11 populations of E. sexmaculatus (Riley).

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    Agricultural Product Processing, Preservation, Storage, Analysis and Detection
    Optimization of Extraction of Procyanidin from Cocoa Bean of Hainan Using Response Surface Methodology
    FANG Yiming,CHU Zhong,GU Fenglin,HE Shuzhen,LAI Jianxiong
    2020, 41 (4):  779-786.  doi: 10.3969/j.issn.1000-2561.2020.04.020
    Abstract ( 593 )   HTML ( 10 )   PDF (8761KB) ( 307 )  

    The extraction of procyanidin from the cocoa beans of Hainan was investigated by the response surface methodology. Single experiments were used to explore the effects of material-to-liquid ratio, extraction concentration, extraction times and extraction temperature on the yield of procyanidin. Based on the analysis of significance and interaction among different factors, the optimal condition of extraction technology was obtained as followed: solvent concentration 67%, material-to-liquid ratio 1∶20 (g/mL), time 88 min and temperature 76 ℃. The extraction rate of procyanidin of the optimized procedure reached 6.21%. The results provide a theoretical basis for the exploitation and utilization of cocoa beans in hainan.

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    Combustion Properties of Rubber Wood Treated with P-N-B Flame Retardants
    JIANG Huichuan,LI Kai,MA Yue,MA Xuemei,HU Zhiyong
    2020, 41 (4):  787-792.  doi: 10.3969/j.issn.1000-2561.2020.04.021
    Abstract ( 631 )   HTML ( 11 )   PDF (1217KB) ( 428 )  

    The influences of different formulations of ammonium dihydrogen phosphate (DAP) and sodium borate (SB) on the combustion properties of rubber wood were studied using the limited oxygen index (LOI) and cone calorimetry test (CONE). The results showed that P-N-B flame retardants (P-N-B) were effective in flame control for rubber wood. When 10.67 wt% of P-N-B was added, LOI increased to 52.3%. Compound flame retardant had a synergistic effect, and the heat release rate, total heat release and total smoke release was reduced by 60%, 50% and 97%, respectively. The combination of DAP and SB could enhance the flame retardancy and smoke suppression property of rubber wood greatly.

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    Chemical Constituents of Stem and Leaf of Three Species of Zingiberaceae Plants in Nujiang, Yunnan Province
    LING Ruimei,LI Ping,LIU Yuanyuan,ZHANG Feifei,YAN Hongbin,WU Lianzhang,YOU Shenjun,YANG Yi,YANG Quan,ZHANG Chunhua,HU Jian,YAN Jian
    2020, 41 (4):  793-803.  doi: 10.3969/j.issn.1000-2561.2020.04.022
    Abstract ( 705 )   HTML ( 7 )   PDF (1296KB) ( 259 )  

    To fully use the stem and leaf of the three species zingiberaceae plants from Nujiang, Yunnan, the chemical composition differences of the stem and leaf of Amomum tsaoko Crevost et Lemarie, Amomum villosum Lour., Alpinia zerumbet (Pers.) Burtt.et Smith were comprehensively evaluated. Six active compounds with anti-cancer, anti- inflammatory, and other activities were analyzed to evaluate the potential pharmacological application. The content of the active compounds in the stem and leaf of the plants was significantly different. Epicatechin was rich in the stem of A. villosum Lour. and A. zerumbet (Pers.) Burtt. et Smith, while heptanediol was rich in the stem of A. tsaoko Crevost et Lemarie, and quinic acid and shikimic acid were rich in the leaf of A. tsaoko and A. zerumbet. There was also a lower content of heptanediol in the leaves of A. tsaoko, much higher epicatechin content of in the leaf of A. zerumbet, while only a lower content of quinic acid in the leaf of A. villosum. The total chemical composition of n-hexane extract was analyzed by GC-MS to find out the composition with potential application value. The results showed that the total chemical composition of the stem and leaf was significantly different in the types and amount. Furthermore, perfume substitutes or precursors with high content were found in the stem and leaf of A. villosum, such as ambria, myrtenol, β-pinene, farnesylacetone, and so on. High content of vitamin E and 5,6-dehydrokavain and other active substances were found in the stem and leaf of A. zerumbet. 1-Hexanoic acid was also found in the leaf of A. tsaoko. This study would provide some references value for the development and utilization of the stem and leaf of the plants.

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    Optimization of Extraction Technology and Stability of Cactus Fruit Pigment
    WEN Chunyan,WANG Caixia,ZHANG Junhao,WANG Chenyu,GUO Dalong,LIAN Haiting,CAO Jun
    2020, 41 (4):  804-810.  doi: 10.3969/j.issn.1000-2561.2020.04.023
    Abstract ( 699 )   HTML ( 7 )   PDF (2594KB) ( 226 )  

    A solvent extraction technique was developed to extract the pigment from the wild cactus fruit in Hainan and optimized using the L9 (3 4) orthogonal test. The stability of the pigment to pH, temperature, light, metal ions and so on was also investigated. The optimal extraction conditions using distilled water were 10 min at pH 7.5 and 30 ℃ with a solid/liquid ratio 1∶20 (g/mL). Under the conditions, the yield of pigment was up to 7.75%. The pigment had good stability under pH 4~10, temperature less than 30 ℃, darkness conditions. The metal ions such as Na+, K+, Ca2+, Mg2+ had no obvious effect on the stability of the pigment. However, metal ions Cu2+, Fe2+, Fe3+, Al3+ could worsen its stability, especially Cu2+ and Fe3+. Oxidizing agent, reducing agent Na2SO3 and Vc reduced the stability of the pigment.

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    Comparative Analysis of Active Ingredients of Different Collection Times from Different Geographical Sources in Anoectochilus roxburghii and Anoectochilus formosanus
    ZENG Biyu,SU Minghua,CHEN Qingxi,LI Huihua,CHANG Qiang,WANG Wei
    2020, 41 (4):  811-818.  doi: 10.3969/j.issn.1000-2561.2020.04.024
    Abstract ( 587 )   HTML ( 7 )   PDF (1407KB) ( 276 )  

    The biomass and active ingredients of four cultivars (‘Sanming NL (narrow leaf)’, ‘Sanming RL (round leaf)’, ‘Zhangzhou NL’ and ‘Taiwan AF’) of Anoectochilus roxburghii and Anoectochilus formosanus from different geographical sources were assayed 120 days after transplantation. The polysaccharides content, fresh weight and dry weight per plant of ‘Zhangzhou NL’ were significantly higher than that of the other three cultivars. Also, the total polyphenols and total flavonoids content of ‘Taiwan AF’ were significantly higher than that of the other three cultivars. The dynamic changes of the active ingredients in different collection periods were investigated. Except for ‘Taiwan AF’ cultivar, the variation rules of total polyphenols, total flavonoids and polysaccharides content in the three varieties were roughly the same. The content of total polyphenols and total flavonoids increased and the content of polysaccharides decreased with the extension of transplanting time. The content of total polyphenols and total flavonoids reached the maximum 150 days after transplantation for ‘Sanming NL’ and ‘Sanming RL’. For ‘Sanming NL’, the content of total polyphenols and total flavonoids was (9.81±0.24) and (4.62±0.16) mg/g, respectively. For ‘Sanming RL’, the content of total polyphenols and total flavonoids were (9.27±0.22) and (4.60±0.17) mg/g, respectively. In ‘Zhangzhou RL’, the content of total polyphenols and total flavonoids reached the maximum 120 days after transplantation. The content of total polyphenols and total flavonoids in ‘Zhangzhou RL’ was (7.74±0.05) and (4.63±0.04) mg/g, respectively. The content of polysaccharides reached the maximum 90, 0 and 120 days after transplantation for ‘Sanming NL’, ‘Sanming RL’ and ‘Zhangzhou RL’, respectively. The content of polysaccharides was (26.05±2.11), (24.29±2.33) and (45.90±4.70) mg/g in ‘Sanming NL’, ‘Sanming RL’ and ‘Zhangzhou RL’, respectively. Besides, the analysis of the active components in different parts in the plant of ‘Sanming NL’ and ‘Sanming RL’ revealed that polyphenols and flavonoids were mainly accumulated in the leaves but sugars in the stems.

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    Analysis on Enzyme Activity and Microbial Community Diversity in Rhizosphere Soil of Different Sugarcane Varieties
    NONG Zemei,SHI Guoying,ZENG Quan,YE Xuelian,QIN Huadong,HU Chunjin
    2020, 41 (4):  819-828.  doi: 10.3969/j.issn.1000-2561.2020.04.025
    Abstract ( 656 )   HTML ( 3 )   PDF (8049KB) ( 285 )  

    In order to provide scientific support for the selection of sugarcane production and the maintenance of sugarcane soil productivity, the differences in nitrogen uptake efficiency of eight sugarcane varieties and the effects of different varieties on the enzyme activities and microbial community diversity in the rhizosphere soil were compared through the barrel cultivation experiment. Sugarcane plants and rhizosphere soils were collected at the end of large growth stage of sugarcane, and nitrogen utilization efficiency, invertase and urease activities in the rhizosphere soil of different sugarcane varieties were measured and compared. Meanwhile, the bacterial community structure of the sugarcane rhizosphere soil was determined and analyzed by the Miseq high-throughput sequencing technology. The nitrogen uptake efficiency of different sugarcane varieties was different. The average nitrogen uptake efficiency of the eight varieties was 17.95%-27.57%, and the nitrogen use efficiency was 22.15%-34.02%. The highest nitrogen uptake efficiency was for GT44, and the lowest was for B9. The invertase activity of the rhizosphere soil was 3.51-6.56 mg/(g·d), and there were significant differences among different cultivars. But the urease activity was 1.12-1.42 mg/(g·d), and there was few differences among different varieties. GT48 and ROC22 were the two varieties with higher soil enzyme activity in the rhizosphere. The results of high-throughput sequencing showed that there were some differences in the bacterial population structure in the rhizosphere soil of the eight sugarcane varieties, but there was no significant difference among different varieties in the composition of the dominant bacteria, and the main difference was in the abundance of the dominant genera. Bacillus was the dominant genus for GT44 and Y93159, while Chryseolinea was the dominant genus for the other six varieties. Redundancy analysis (RDA) showed that the nitrogen absorption efficiency of sugarcane, the content of available N and pH value of soil had the greatest effect on the soil microbial community structure. Correlation analysis showed that the nitrogen absorption efficiency of sugarcane and soil physiochemical characteristics were significantly correlated with different dominant microbial communities. This study would deepen the understanding of the microbial community in the sugarcane rhizosphere and provide references for the relation between the microbial composition and diversity with environmental factors.

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    Soil Biological Properties in Rhizosphere of Pea (Pisum sativum L.) Improved by Magnetic Treatments
    ZHANG Chuanjin,REN Kuiyu,GUO Shuang,PANG Shichan,WANG Shuaishuai,YANG Shangdong
    2020, 41 (4):  829-836.  doi: 10.3969/j.issn.1000-2561.2020.04.026
    Abstract ( 576 )   HTML ( 2 )   PDF (1760KB) ( 257 )  

    To explore the effect of magnetic treatments on the soil fertility and health, the soil biological properties in the rhizosphere of peas were analyzed using pot culture. Five treatments, i.e. No-treatment (CK), iron powder (A), magnetized iron powder (B), magnetized iron powder+0.2 T magnet (C) and 0.2 T magnet treatments (D) were set for analyzing with the high-throughput sequencing technique. The amounts of cultivable bacterial and fungi could be significantly increased by treatment C. Meanwhile, compared to CK, the biomass C, N, P and the activities of β-Glucosidase, Aminopeptidase and Phosphatase were also increased by 34.0%, 46.0%, 44.2% and 24.6%, 23.7%, 31.8% compared to treatment C, respectively. In addition, the indexes of richness and diversity of bacteria, such as Ace, Chao1 and Shannon in treatment C were also significantly higher than those in CK. Moreover, Sneathiella, Desulfurispora and nannocystaceae were the unique bacteria in CK under genus level, but seven different bacterial groups under genus level were showed in treatments A and B. Eleven different bacterial groups were presented in treatment C. And there were only eight different bacterial groups in treatment D. In summary, the biological indicators of soil fertility and health, such as soil cultivable microorganisms, microbial biomass, enzyme activities and bacteria diversity were all significantly improved under treatment C, which used 0.2 T magnet + 5 g magnetized iron powder. It indicated that the soil fertility and ability to resist soil-borne diseases could be promoted by treatment.

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    Evaluation of the Potentiality of Asteraceae Flowers as Green Manure in Summer Pear Orchard
    YAO Danjun,ZHANG Aihua,WU Xinghong,RAN Bin,ZHANG Qin,ZHU Qing,WANG Wenhua,CAO Weidong,KUANG Shengjian
    2020, 41 (4):  837-844.  doi: 10.3969/j.issn.1000-2561.2020.04.027
    Abstract ( 533 )   HTML ( 2 )   PDF (1472KB) ( 189 )  

    Asteraceae flowers are widely used in various landscapes. The research aims to explore the potential of green manure utilization by planting different varieties of Asteraceae flowers in pear orchards, and choose out better varieties to provide theory and practice for the sustainable development of tourism agriculture. Seven common summer Asteraceae flowers were used as the experimental materials. The potential of green manure utilization was obtained by comparing the content and accumulation of nutrients and the effects on soil nutrients. The growth rate of different species of Asteraceae in different growth stages was different. The plant height and root length of oil sunflower were the highest at the blossom stage, which was (193.17±20.71) and (18.43±3.72)cm, respectively. The flower traits and flower blossom time of different flowers were different, but the inflorescence area could completely cover the ground during the blossom period, which had a good ornamental value. Among them, the inflorescence area of oil sunflower was the largest, and the average inflorescence was about (0.088±0.010)m2, and zinnia had the longest flowering period of 107 days. All varieties of Asteraceae flowers could accumulate nutrients in growth. The content of nitrogen, phosphorus and potassium of the aboveground part of zinnia was the highest, which was (2.89±0.07)%, (0.53±0.03)% and (3.27±0.17)% respectively. The dry matter quality, nitrogen, phosphorus and potassium nutrient accumulation and carbon sequestration per unit area of oil sunflower were significantly higher than that of other treatments, which was (18821.17±568.91) (396.89±22.74), (57.35±5.25) and (385.20±22.09)kg/hm2, (2820.96±84.49)g/m2, respectively. Varieties of Asteraceae flowers could affect the soil fertility, and significantly improve soil organic matter, fertilize soil, and increase soil available nitrogen, available phosphorus and available potassium. Based on the comprehensive fertilizer efficiency indicators and the convenience of operation, the sequence of green manure utilization potential of seven species of Asteraceae in pear orchards was: small sunflower > high-pole cosmos > zinnia > dwarf cosmos > high-pole sulphur > dwarf sulphur > oil sunflower. Therefore, taking into account both the beauty and ecology, small sunflower, high-pole cosmos and zinnia could be plantedin the pear orchards.

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