Acta Agriculturae Zhejiangensis

Study on nitrogen nutrition diagnosis of rice leaves based on hyperspectrum

YANG Hongyun, ZHOU Qiong, YANG Jun, SUN Yuting, LU Yan, YIN Hua

2019, 31(10): 1575-1582. DOI: 10.3969/j.issn.1004-1524.2019.10.01

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In order to realize rice nitrogen nutrition diagnosis quickly and accurately, rice cultivation experiments with 4 nitrogen application levels were conducted on Zhongjiazao 17 rice cultivar. A total of 240 rice spectral data were collected at tillering stage within 350 to 2 500 nm from top trilobate by using portable earth mass spectrometry. All the samples were randomly divided into training set (160 samples) and prediction set (80 samples). The original spectrum was pretreated by multiplicative scatter correction (MSC), standard normal variate (SNV) and Savitzky-Golay smoothing (SG) methods, respectively. Then, principal component analysis (PCA) and successive projection algorithm (SPA) were used for feature reduction and feature selection of the pre-processed spectra. After principal component analysis, the first 24 principal components, of which the accumulative contribution exceeded 99.98%, were used as the input variables of the model, and 12, 15 and 19 characteristic wavelengths were selected for the spectrum after MSC, SNV and SG treatments, respectively. Finally, rice nitrogen nutrition diagnosis models were established with support vector machine (SVM). It was shown that the MSC-PCA-SVM model was the best method for rice nitrogen nutrition diagnosis, of which the accuracy rate on training set and prediction set was 99.38% and 97.50%, respectively.

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Effects of spraying boron and molybdenum fertilizers on productivity and nutritional quality of alfalfa under drip irrigation

MIAO Xiaorong, LIU Junying, ZHANG Qianbing, LI Feifei, SUN Yanmei, YU Lei, MA Chunhui

2019, 31(10): 1583-1590. DOI: 10.3969/j.issn.1004-1524.2019.10.02

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To study the effects of spraying boron and molybdenum fertilizers on hay yield and nutritional quality of alfalfa under drip irrigation in oasis region of Xinjiang, four treatments, i.e., application of molybdenum (Mo), application of boron (B), application of boron and molybdenum (B+Mo), and without application of fertilization (CK) were established. Growth traits, hay yield, and the nutritional quality of alfalfa were studied. It was shown that the highest total hay yield of alfalfa was 17 186.41 kg·hm^-2 under B+Mo. There was no significant difference on total hay yield of alfalfa between B and B+Mo. Among indexes of plant height, stem diameter, leaf-stem ratio, growth rate, the highest correlation with hay yield was found in plant height, yet the lowest correlation was found in leaf-stem ratio. The crude protein content of alfalfa under B+Mo was increased by 13.96%-27.64% than that of CK. The crude protein content of alfalfa under B+Mo and B was significantly (P<0.05) greater than that under CK except the third cut, while the crude protein content of alfalfa under B+Mo was significantly (P<0.05) greater than that under Mo except the fourth cut. The acid detergent fiber (ADF) and neutral detergent fiber (NDF) contents of alfalfa under B+Mo were significantly (P<0.05) lower than those under CK. The total hay yield of alfalfa was significantly (P<0.01) positively correlated with crude protein content and relative feeding value. The crude protein content of alfalfa was significantly (P<0.05) positively correlated with the relative feeding value, and the ADF content of alfalfa was significantly (P<0.05) positively correlated with the NDF content. It was concluded that application of B and Mo was beneficial to promote the formation of alfalfa hay yield and to improve the nutritional quality under drip irrigation in Xinjiang oasis area, and the effect of application of B on alfalfa under drip irrigation was better than that of application of Mo.

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Changes of rumen bacterial flora after starch induced milk fat depression in dairy cows

XU Xiaofeng, GUO Cheng

2019, 31(10): 1591-1598. DOI: 10.3969/j.issn.1004-1524.2019.10.03

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This experiment aimed to study the effects of starch supplementation on milk fat and rumen bacterial flora of dairy cows. Four Chinese Holstein cows with similar body conditions and healthy lactation were selected. At the first stage of 7 days, cows were fed by basic diet to be control group. After the second stage of 15 days, cows were fed by basic diet supplemented with an additional 1.5 kg of corn starch per day per head to be experimental group. The results showed that milk fat percentage of dairy cows decreased significantly after starch supplementation (P<0.01). At the phylum level, 28 phyla of rumen bacteria were detected, the richness of Gemmatimonadetes and Tenericutes decreased significantly (P<0.01), Armatimonadetes, Acidobacteria and Cyanobacteria decreased significantly (P<0.05), Firmicutes increased significantly (P<0.05). At the level of genus, 383 genera were detected, 90 of which changed significantly. At the genus level, the abundance of Acetobacter, Lactobacillus, Pediococcus, Leuconostoc, Lachnoclostridium-1, Ruminococcus-1 and Prevotella-7 increased significantly (P<0.01), Anaerovibrio, Enterococcus and Enhydrobacter decreased significantly (P<0.01). In the experiment, flora diversity and distribution uniformity of rumen bacteria in daily cows were not different after adding starch in diet, while proliferation of most acid-producing bacteria was promoted.

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Molecular typing, biofilm formation ability and drug resistance of pathogenic Escherichia coli isolated from pigs in Sichuan Province

PENG Kenan, ZHOU Xueke, YIN Xinhuan, LI fei, CAI Yao, ZENG Yubing, JIANG Chaoyuan, ZHANG Rubo, YANG Zexiao, XU Zhiwen, ZHU Ling

2019, 31(10): 1599-1607. DOI: 10.3969/j.issn.1004-1524.2019.10.04

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The experiment was to study the molecular characteristics and epidemic trends of pathogenic Escherichia coli from pigs in Sichuan Province. From 2016 to 2018, 208 samples of liver, spleen, lung, and intestines of sick pigs were collected from various parts of Sichuan Province. 87 strains of Escherichia coli were screened by bacterial isolation and identification methods, and 22 strains of pathogenic Escherichia coli were determined by SPF mouse challenge test. Molecular grouping, biofilm formation ability detection, pathogenicity and drug resistance of the isolates were studied, drug resistance genes and three HPI virulence genes of pathogenic Escherichia coli were detected by PCR. The results showed that 22 pathogenic Escherichia coli were distributed in group A (59.09%), the positive rate of biofilm formation was 72.73%, and the strong film-forming ability strain accounted for 40.91%. There are different degrees of resistance to 19 antibiotics, among which nine drugs such as enrofloxacin, compound sulfamethoxazole, ampicillin, amoxicillin, erythromycin and tetracycline had higher resistance rates (81.82%-100%). The resistance rates of gentamicin, levofloxacin, norfloxacin and ceftriaxone were between 63.64% and 72.73%. The resistance rate to other drugs was between 4.55% and 54.55%, and all of the 22 isolates were multi-drug resistant. Twelve drug resistance genes detected by 22 pathogenic Escherichia coli include β-lactam TEM(36.36%), SHV(9.09%), OXA(9.09%), sulfonamide sul Ⅰ(63.64%), sul Ⅱ(95.45%), aminoglycosides acc(3)-Ⅱ(68.18%), aph(3')-Ⅱ(50.00%), chloramphenicol cmlA (45.45%), floR(81.82%), tetracycline tetA(81.82%), tetB(86.36%), quinolone qurB(40.91%). Three HPI virulence genes irp1 (40.91%), irp2 (13.64%), and fyuA(13.64%) were detected. It indicated that the pathogenic Escherichia coli from pigs in Sichuan Province was mostly distributed in group A, the positive rate of biofilm formation was high and there was serious multi-drug resistance. The carrying rate of tetracyclines, chloramphenicols, aminoglycosides and sulfonamide-resistant genes were high, and there were multiple drug resistance genes and virulence genes coexisting. The results can provide reference for the prevention and treatment of pig colibacillosis in Sichuan Province.

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Association analysis between polymorphism of ESR1 gene and egg quality in quail

BAI Junyan, FAN Hongdeng, CAO Heng, PANG Youzhi, SONG Xiaojie, JIANG Mengjuan, FU Xueyan, LU Xiaoning, YANG Shuai, LI Xinyue, HAO Weiguang, LI Ziheng, ZHENG Feiyang

2019, 31(10): 1608-1614. DOI: 10.3969/j.issn.1004-1524.2019.10.05

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In order to investigate the relationship between the polymorphism of estrogen receptor (ESR1) gene and quail egg quality, the polymorphism of ESR1 gene was detected by PCR-RFLP in three populations, including Chinese yellow quail, Korean quail and Beijing white quail, and the correlation between the polymorphism of ESR1 gene and quail egg quality traits was analyzed. The results showed that CC, CT and TT genotypes were detected in exon 1, exon 4 and exon 8 of quail ESR1 gene in Beijing white quail, Chinese yellow quail and Korean quail. Exon 1 showed the highest frequency of CC genotype in all three quail populations, Exon 8 showed the highest frequency of TT genotype in all three quail populations, Exon 4 showed the highest frequency of TT genotype in Chinese yellow quail and Korean quail, 0.409 and 0.617, respectively, while Beijing white quail had the highest frequency of CC genotype (0.667).The results of correlation analysis showed that the yolk height and yolk index of CC genotype and CT genotype in exon 4 were significantly higher than TT genotype (P<0.05), the yolk width of TT genotype was significantly higher than CC genotype and CT genotype (P<0.05), and the shell thickness of CT genotype and TT genotype was significantly higher than CC genotype (P<0.05). In exon 8, the yolk index of CC genotype was significantly higher than that of CT genotype and TT genotype (P<0.05), and the yolk width of TT genotype of CT genotype was significantly higher than that of CC genotype (P<0.05). ESR1 gene can be used as a valuable candidate gene for molecular marker-assisted selection in egg quails.

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Universal primer screening and verification for fish environmental DNA research based on high-throughout sequencing technology

ZHANG Aiju, HAO Yabin, GUO Aihuan, LIU Jindian, LIAN Qingping, ZHOU Zhiming

2019, 31(10): 1615-1623. DOI: 10.3969/j.issn.1004-1524.2019.04.06

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In order to screen one or more applicably universal primers for environmental DNA(eDNA) research based on high-throughout sequencing technology, five pairs of primers of fish mitochondrial genome fragment were chosen from literatures, which could amplify the partial sequences of Cyt b gene, 16S rDNA and COⅠ gene. After initial amplification on two water eDNA samples, it was found that CY45M and 1634M had better amplification effects and their products could be applied to the subsequent high-throughout sequencing. Meanwhile, no purpose band could be amplified using the other three primers, including CYFM, 16ACM and COⅠM. After the next amplification on 7 water eDNA samples using CY45M and 1634M, it was found that all the eDNA were amplified by these two pairs of primers, and the PCR products presented clear bright bands by gel electrophoresis. According to the high-throughout sequencing and BLAST results, the sequences gained by both of CY45M and 1634M could be annotated on 11 fish species.Nevertheless, there were dissimilarity and repeatability upon these fish species. In summary, the CY45M and 1634M were both suitable universal primers for study on eDNA of fish community structure in Qiantang River.

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Genetic diversity analysis and fingerprinting construction of Muscat Hamburg grapevine series by iPBS marker

ZHANG Anshi, PANG Jincheng, LIU Ying, GAO Dengtao, WEI Zhifeng

2019, 31(10): 1624-1631. DOI: 10.3969/j.issn.1004-1524.2019.10.07

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In order to explore the genetic relationship among the germplasm resources of Muscat Hamburg grapevine series, the genetic diversity of 39 cultivars was analyzed by iPBS marker, and their DNA fingerprints were constructed. 14 primers were screened for PCR amplification of 39 cultivars. A total of 152 bands were generated, of which 132 bands were polymorphic bands (86.86%).The average value of polymorphism information content, number of observed alleles, number of effective alleles, Nei's gene diversity index and Shannon's information index were 0.863 8, 1.868 4, 1.409 0, 0.251 8 and 0.390 7, respectively. The genetic similarity coefficient among 39 cultivars ranged from 0.526 3 to 0.940 8 with a range of 0.413 5. The above results indicated that there were rich genetic diversity among 39 cultivars. The dendrogram of these 39 cultivars was constructed using UPGMA. They could be divided into 4 groups at genetic similarity coefficient of 0.72, of which V. vinifera were mainly distributed in Group Ⅰ, while the Euro-American hybrids species were mainly distributed in Groups Ⅱ, Ⅲ and Ⅳ. The DNA fingerprints of 39 cultivars were constructed using 16 polymorphic loci amplified of 8 primers, providing scientific basis for the identification of Muscat Hamburg grapevine series.

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Effects of rain-sheltered cultivation on fruit quality in mid-late maturing juicy peach

XIAO Jinping, CHANG Luwei, ZHANG Huiqin, XIE Ming, GU Xianbin, XIONG Caizhen, WU Dajun

2019, 31(10): 1632-1638. DOI: 10.3969/j.issn.1004-1524.2019.10.08

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Rain-sheltered cultivation is an important technical measure to promote peach industry in southern China. In this study, three mid-late maturing peach cultivars, Xiahui No.5, Danxiayulu and Chiyulu were used as test materials to analyze the influence of rain-sheltered cultivation on fruit quality of peach cultivars by using open-field cultivation as the control. The results showed that the rain-sheltered cultivation increased the soluble solid content, solid-acid ratio and sucrose content in the fruits, and especially for late-maturing cultivar Chiyulu, the content of sucrose, fructose and glucose of fruits were significantly higher than that under open-field cultivation. Overall, the contents of total acid and organic acid in fruits of mid-maturing cultivars showed a downward trend after rain-sheltered cultivation, while that of the late-maturing cultivar Chiyulu was the opposite. The rain-sheltered cultivation also increased the content of soluble sugar, soluble protein and anthocyanin in the peels, meanwhile, the contents of total phenols and flavonoids of the peels reduced. In addition, the rain-sheltered cultivation also caused the average fruit weight and color parameters of some cultivars to decrease obviously. Therefore, compared with the open-field cultivation, the rain-sheltered cultivation could significantly improve the flavor quality of the peach fruits, but at the same time it also had some adverse effects on the yield and appearance coloring of the fruits to some extent. Rain-sheltered cultivation techniques should be optimized and adjusted in full consideration of cultivar characteristics and specific weather conditions.

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Effects of summer pruning on fruit quality and 6 kinds of aromatic compounds of Jumeigui grape

SHANG Jiayin, LI Kai, WANG Chaoxia, SUN Jianjun, JI Xian, WANG Dan, SU Hong, ZHANG Xiaolei, TIAN Shufen

2019, 31(10): 1639-1646. DOI: 10.3969/j.issn.1004-1524.2019.10.09

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In order to explore the effects of different summer pruning methods on the fruit development, quality, alcohol and aldehyde compounds content of Jumeigui grape, the leaf-to-fruit ratio during the grape growing season was set to 20∶1, and the “three-stage” pruning method was used to be set 3 kinds of pruning methods which was “4+8+8”, “4+12+4” and “4+4+12”. The weight, volume, total soluble solid (TSS) content, procyanidine, flavonoids, total phenols, hexanal, 2-hexenal, hexanol, 2-hexene-1 alcohol, nerol, linalool contents, and other indicators of grape fruit were measured once every 2 weeks from 8 weeks after flowering. Correlation analysis was used to analyze the correlation between the indicators. The results showed that treatment B (“4+12+4”) was superior to the other two treatments in fruit development, TSS and the content of 1-hexanol, nerol and linalool. The fruit quality of the “three-stage” pruning method which left relatively more leaves in the middle of the new shoots was better than that of the less leaves. The content of 1-hexanol, nerol and linalool were significantly or highly significantly positively correlated with the grape ripening index which were single weight, single volume, soluble solids and proanthocyanidin content. The increasing strength of the content of nerol and linalool was the largest before and after fruit maturation period. In summary, the summer pruning method could affect the quality of the grape fruit, and the summer pruning method of treatment “4+12+4” was the best of all.

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Comparative study on photosynthetic characteristics and yield of different switchgrass varieties in saline-alkali land of northwest China

CHANG Wenwen, LIU Jili, WU Na, ZHANG Yongqian, HE Haifeng

2019, 31(10): 1647-1654. DOI: 10.3969/j.issn.1004-1524.2019.10.10

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To screen switchgrass varieties with strong photosynthetic performance and high yield, eleven varieties of energy crop-switchgrass were used as materials in Yinbei saline-alkali region of Ningxia, using grey correlation analysis method to study yield and five photosynthetic indexes which were transpiration rate, stomatal conductance, net photosynthetic rate, intercellular CO₂ concentration, water use efficiency. The results showed that there were significant differences in photosynthetic characteristics and yield among different switchgrass varieties at flowering stage under saline-alkali soil conditions. The general performance were Cave-in-Rock>Alamo>Kanlow>Forestberg>Ansai>Pathfinder>Nebraska 28>Japan>Trailblazer>NewYork>Black Well. Among them, Cave-in-Rock had high yield and the highest weighted correlation degree with ideal varieties, comprehensive performance was also the best. The weighted correlation degrees of Alamo and Kanlow were 0.711 and 0.654, and their biomass yield were 20.47 and 17.93 t·hm^-2, respectively. Comprehensive evaluation showed that Cave-in-Rock, Alamo and Kanlow had better potential yield than the others in saline-alkali region. Black Well, NewYork, Trailblazer, Japan and Nebraska 28 had lower yield of biomass and photosynthetic characteristics, while the comprehensive performance of other varieties were medium level.

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Identification and culture conditions of a wild-type Termitomyces aurantiacus strain from Lincang

ZHOU Huiming, ZHANG Yanzhen, CHAI Hongmei, YANG Rongqing, TAN Ying, ZHANG Pingping, BAI Yuying, ZHAO Yilian, JIN Yujie

2019, 31(10): 1655-1662. DOI: 10.3969/j.issn.1004-1524.2019.10.11

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To screen the optimal culture condition for mycelial growth of Termitomyces aurantiacus and lay the foundation for its bionic cultivation, strain LCT10 was isolated from a wild mushroom fruiting body of Termitomyces from Lincang, Yunnan. Taxonomic status was identified based on its morphology and ITS sequence analysis, and the different culture conditions were designed to determine the effect of physical factors (temperature, light intensity and aeration) and chemical factors (pH, carbon resources, nitrogen sources and C/N) on mycelial growth of T. aurantiacus. The result showed that the strain LCT10 was T. aurantiacus, its morphological characteristics were consistent with its molecular identification. Its optimal temperature and pH were 25 ℃ and 8.0. It was able to grow in 5 kinds of ventilation treatments, and the growth of mycelia was the fastest in the treatment of no ventilation. Soluble starch, corn flour, sucrose, honey and glucose were suitable for its growth, honey was the best carbon soure for mycelial growth. Peptone, glycine, ammonium nitrate and urea could be used as nitrogen sources and peptone was the best. The optimum C/N for mycelial growth was (20-30)∶1. The growth of strain LCT10 under the best culture conditions had significant difference at level of 0.05 comparing with other treatments

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Cloning of sublitisin-like protease gene cDNA from Isaria cicadae and characterization of its sequence and deduced protein

CHEN Guanju, CHAI Yiqiu, LI Xiaola, FANG Ming, LIU Yougao, JIN Yiwei

2019, 31(10): 1663-1670. DOI: 10.3969/j.issn.1004-1524.2019.10.12

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Isaria cicadae is one of the most common species of entomopathogenic fungi, and has been used as biocontrol agents against pests. In order to obtain a complete cDNA of sublitisin-like protease from I. cicadae, a pair of primers was designed using MegAign of DNAStar analysis software according to the high conserved nucleotide sequences in GenBank, and RT-PCR, 3'/5'-RACE PCR were used. The whole sequence of cloned cDNA was 2 031 bp, and the results showed that it had 170 bp and 262 bp on the 5'-terminal and 3'-terminal respectively, encompassed an open reading frame (ORF) with 1 599 bp encoding 532 amino acid (aa) residues, including 18-aa signal peptide, 133-aa propeptide. Alignments with the deduced amino acid of proteins in the species of Isaria farinose, Beauveria bassiana, Cordyceps militaris and Trichoderma harzianum showed a homology of 88%, 88%, 89% and 71%, respectively. The sublitisin-like protease of I. cicadae was located in the vacuoles of cells associated with the secretory pathway, with 5 cycteines and 4 putative N-glycosylation sites. The secondary structure of the protein was mainly the α-helix and randon coil.

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Screening, identification, and characteristics of an antagonistic strain against Fusarium oxysporum f.sp. niveum

WANG Hengxu, XU Weihui, YANG Youcai, WANG Zhigang, LIU Zeping, WANG Kexin

2019, 31(10): 1671-1680. DOI: 10.3969/j.issn.1004-1524.2019.10.13

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In order to screen an antagonistic strain against Fusarium oxysporum f.sp. niveum (Fon), the rhizosphere soil from vegetable crops on Qingxin farm in Qiqihar City was collected and used in this study. The antagonistic strain was screened out using plate confrontation method, and identified by morphological, 16S rDNA, gyrA and gyrB sequence analysis. Pot experiment was conducted to verify the control effect of antagonistic strains on Fusarium wilt of watermelon. Growth rate method was used to study antifungal activities of sterile fermentation filtrate from different growth stages of antagonistic strain and the stability of sterile fermentation filtrate. Fon conidia morphology and cell membrane integrity were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), respectively. The results showed that the antagonistic strain WD was Bacillus amyloliquefaciens, which could effectively inhibit the occurrence of watermelon wilt, and the inhibition rate reached 57.57%. The sterile fermentation filtrate had higher antifungal ability in the decline period, and the growth inhibition rate of Fon mycelium was 54.32%. Sterile fermentation filtrate from B. amyloliquefaciens WD had high tolerance to temperature and pH. The antifungal activity was the strongest at 30 °C and pH 7, the inhibition rate reached 58.36%; The sterile fermentation filtrate could store at 4 °C for at least 45 days, and UV light for 120 min had no effect on its antifungal activity. The Fon conidia were exposed to the sterile fermentation filtrate from B. amyloliquefaciens WD, the surface of the conidia collapsed and the membrane integrity was destroyed. In conclusion, B. amyloliquefaciens WD had high-efficiency antifungal activity on Fon, the results laid a foundation for development and utilization of biocontrol agent to Fusarium wilt of watermelon.

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Conversion dynamics and effectiveness of ammonium polyphosphate with different polymerization degrees to soil phosphorus

CHEN Xiaojuan, CHEN Yulin, LIN Jingjing, YANG Yibin, HU Kewei, ZHANG Chenglin

2019, 31(10): 1681-1688. DOI: 10.3969/j.issn.1004-1524.2019.10.14

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In the present study, calcareous soil and laterite soil were selected as test materials, and soil culture experiment was carried out to explore the effect of ammonium polyphosphate (APP) with different polymerization degrees on the dynamic transformation and availability of soil phosphorus. Five treatments were set up: CK, no phosphate fertilizer; MAP, application of monoammonium phosphate; APP_L, application of low polymerization degree APP; APP_M, application of medium polymerization degree APP; APP_H, application of high polymerization degree APP. It was shown that, in calcareous soils, the available phosphorus content under APP_H was significantly (P<0.05) lower than that under MAP at 5 d, yet it was significantly (P<0.05) higher than that under MAP by 23.92% at 50 d. In laterite soil, the available phosphorus content under APP_H was significantly (P<0.05) lower than that under MAP in 5-10 d, yet it was significantly (P<0.05) higher than that under MAP in 15-50 d. The available phosphorus content under APP_L was significantly (P<0.05) higher than that of MAP in 5-15 d, yet there was no significant difference between the two treatments in 30-50 d in laterite soil. In calcareous soils, the content of Resin-P under APP_L and APP_H was higher than that under MAP during the whole experiment, while the content of NaCHO₃-P under APP_H increased with the prolongation of culture time, which was higher than that under other treatments. In laterite soil, the contents of Resin-P and NaHCO₃-P under APP_L and APP_H were lower than those under MAP initially, yet they got higher than those under MAP with the increasing culture time. In calcareous soil, APP_L and APP_H could increase the availability of soil phosphorus as compared with MAP, and reduce phosphorus fixation. In laterite soil, APP_L exhibited high availability of phosphorus in the early stage, which was suitable for basal application, yet APP_H showed high availability of phosphorus in the later stage, and reduced phosphorus fixation in acidic soils.

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Diversity of ancient tea germplasm based on morphological characteristics in Guizhou

NIU Suzhen, SONG Qinfei, AN Hongwei, HUANG Zheng, LIU Xia, ZHAO Degang, CHEN Zhengwu

2019, 31(10): 1689-1699. DOI: 10.3969/j.issn.1004-1524.2019.10.15

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In this study, 144 ancient tea germplasm distributed in 32 zones were evaluated to analyze the variation of morphological characters and the diversity, and to provide theoretical basis for the utilization of ancient tea germplasm by morphological way in Guizhou. The results showed that the variation coefficients of 9 morphological traits were above 35% except tree posture, and the diversity index of 10 morphological traits was above 0.85; the cumulative contribution rate of the first four principal component factors was only 85.97%. Among the five ecological zones, the morphological and genetic diversity of ancient tea germplasm resources in southwestern Guizhou was wider than that in other zones. Three groups were divided at the distance of 5.60. The first group included 27 samples, which mainly came from southeastern Guizhou, central Guizhou, southwestern Guizhou and northern Guizhou. The second group included 10 samples, which mainly came from Qinglong in southwestern Guizhou. The third group included 107 samples, which were widely distributed.

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Simultaneous detection and analysis of 16 kinds of mycotoxins in Pu-erh tea

HU Lin, SHI Zhen, ZHAO Li, LIU Xiaosong, DONG Yuying, JIANG Mengyuan, LI Wenting

2019, 31(10): 1700-1708. DOI: 10.3969/j.issn.1004-1524.2019.10.16

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To establish a convenient and accurate method for the determination of 16 mycotoxins with liquid chromatography-tandem mass spectrometry(LC-MS/MS) in Yunnan Pu-erh tea, including aflatoxin, fumonisin (FB₁-FB₂), nivalenol, deoxynivalenol (DON), zearalenone, ochratoxin A, T-2 toxin, HT-2 toxin and heterochromatoxin, samples were extracted by organic solvent (methanol-water-formic acid volume ratio was 70∶29∶1), purified by PriboFast^® Multi-Toxin IAC immunoaffinity purification column, determined and analyzed by multi-reaction monitoring of LC-MS/MS, and quantified by internal standard method. The regression equations of 16 mycotoxins had a good linear relationship(r>0.99). The recovery rate at three different levels was 84.2%-102.7%, and the relative standard deviation was 2.18%-4.78%. In the detection of 16 kinds of mycotoxins in 174 samples of Yunnan Pu'er tea, DON was detected in 53 samples of tea with the detection rate of 30.63%, FB₁ was detected in 96 samples of tea with the detection rate of 55.17%, and FB₂ was detected in 12 samples of tea with the detection rate of 6.90%, FB₃ was detected in 12 samples of tea with the detection rate of 6.90%. Other mycotoxins were not detected. The contents of the 4 mycotoxins detected were between 0.2-6.0 μg·kg^-1, which did not exceed the international food safety limit. The detection rate of mycotoxins in Pu-erh tea in Yunnan Province was low, and the content of mycotoxins met the requirements of national standards. The method was accurate, reliable and convenient, which can be applied to the detection of mycotoxins in tea, especially in batch tea.

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Self-alignment algorithm of wheat plot breeding path based on Beidou satellite positioning

WANG Hao, TANG Yongwei, DONG Zhenzhen, WANG Maoli, ZHAO Jingbo

2019, 31(10): 1709-1716. DOI: 10.3969/j.issn.1004-1524.2019.10.17

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Breeding experts need to accurately divide the sowing areas in wheat plot breeding. However, in the sowing process of the traditional wheat plot planter, the lines in each plot cannot be aligned precisely, leading to confusion in the breeding area and affecting the breeding effect. In order to solve the problem of precise alignment of the planting path in wheat plots, Beidou satellite positioning was combined with wheat plot breeding on the basis of long-term experiments. Firstly, the longitude and latitude of the planter was obtained, and the plot path model of the planter was built through the establishment of relative coordinate system to obtain the planned path, which replaced the traditional manual line drawing process. Using the method proposed in this paper, the sowing accuracy of wheat plot was increased by 32% within the error range of 5 cm. At the same time, considering the complexity of farmland operation environment and limited ability of manual driving, Beidou satellite positioning itself also had centimeter-level error. In order to further improve the accuracy of path planning, this paper put forward a kind of arithmetic average value filtering and the algorithm of Kalman filtering fusion. The fused algorithm performed iterative recursive processing on the slope value of the reference path in the relative coordinate system. Through experiments, the arithmetic mean filtering and Kalman filtering combined algorithm could effectively correct the slope value of the actual calculation, with the minimum correction range of 20.1% and the maximum correction range of 4 871.4%. The research results in this paper could provide reference for the accurate planning of the Beidou satellite positioning in the planting path.

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Study on general models for non-destructive inspection of leaf moisture content of 10 plants

ZHENG Junbo

2019, 31(10): 1717-1723. DOI: 10.3969/j.issn.1004-1524.2019.10.18

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Rapid, precise and non-destructive determination on moisture content of plant leaves have contributed to diagnosis of water deficiency. Ten plants were used as samples to detect their leaf capacitance and resistance by using the self-designed parallel-plate capacitor and improving resistance measuring method. Measured data was analyzed by SPSS 19.0 software for intraclass correlation coefficient (ICC) to verify reliability of the data. The leaves were divided into training set and test set. The training set was analyzed with Excel regression. The fitting equation was established among leaf moisture content, capacitance and resistance. The fitting equation was used to predict the leaf moisture content in the test set. It was shown that the data reliability of leaf capacitance among 10 plants was good. The data reliability of leaf resistance was good in Photinia × fraseri Dress and Myrica rubra (Lour.) S. et Zucc., and general in Ligustrum lucidum Ait., Sapindus mukorossi Gaertn, Cercis chinensis and Osmanthus sp., and poor in Viburnum odoratissinum. By Excel regression analysis, coefficient of determination (R²) was 0.978 8, adjusted R² was 0.977 4, significant value P=7.85×10^-37, fitting equation Z=86.0897-628.471X^-1-11.1753Y+117.2954Y·X^-1, the fitting effect of the model was good. The errors were -2.53%-1.46% compared with the drying method to predict the moisture content of test set with this model. This model could be used as a generic model to predict leaf moisture content of these 10 plants.

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Analysis of benefit-linking mechanism of farmers' cooperatives in industrial convergence: based on research of three state-level model cooperatives

SUN Kun, FANG Keyu, TONG Shanshan, ZHANG Shemei

2019, 31(10): 1724-1733. DOI: 10.3969/j.issn.1004-1524.2019.10.19

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In the present study, three state-level model cooperatives were selected as examples to analyze the benefit-linking mechanism of cooperatives in promoting the integration of primary, secondary and tertiary industries from four aspects of interest generation, interest distribution, incentive system and risk prevention and control, and the commonness and differences of various benefit-linking types were discussed. It was shown that the cooperatives must consider establishing different interests with internal members and external capital (“one cooperative, two systems”) to promote the industrial convergence. The interest distribution system should be innovated in combination with cooperation background, trust mechanism and other factors to stabilize the interests of members. Cooperatives should pay attention to the attraction of profitable projects and the appeal of external capital interests when introducing external capital, and ensure the dominant position of cooperatives. Therefore, to promote the industrial convergence, cooperatives should promote the new idea of “one cooperative, two systems”, accurately promote the stability and upgrading of members' interests, and build a new pattern of win-win cooperation featuring equality and mutual benefit.

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Whole chain closed-loop pattern on development and utilization of livestock and poultry germplasm resources: an illustration from Yukou Poultry Co., Ltd.

CHENG Hua, LU Fengjun, LIU Qing

2019, 31(10): 1734-1744. DOI: 10.3969/j.issn.1004-1524.2019.10.20

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The present paper summarized the fundamental problems faced by the development and utilization of livestock and poultry germplasm resources (LPGR) by means of enterprise investigation and field interview, and analyzed the scientific and technological innovation, service and organization mode of Yukou Poultry Co., Ltd. in developing and utilizing LPGR by means of literature review and systematic analysis. It was shown that the varieties innovated on LPGR could be developed to good seed with market consumption value identification as the fundamental guide, along with the consideration of the performance of breeding, production, commodity with the closed whole industry chain. Service complex could effectively promote the transformation of new varieties, with the fundamental goal of serving the whole life process of innovative varieties. “Leading enterprises-services platform-culture cluster” platform-oriented industrial organization model could effectively promote the development and utilization of LPGR, with “own and united” and “self-construction and join” forming the scale and scope economy of service, breeding and the transformation of innovation varieties. In order to realize the virtuous circle of the value between the development and innovation of LPGR and the transformation, the main bodies in the industrial chain should develop the joint consciousness to form the platform organization ecology with comprehensive service, and the government should support the leading subjects to carry out the construction of systematic development and utilization ability based on the full chain.

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Effect of health consciousness on pro-environmental behavior of scale breeding farmers: based on payment willingness for environmentally friendly feed

WANG Anbang, HE Ke, ZHANG Junbiao

2019, 31(10): 1745-1757. DOI: 10.3969/j.issn.1004-1524.2019.10.21

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In order to explore the effect of health consciousness on the pro-environmental behavior willingness of large-scale pig farmers represented by environmentally friendly feed, 402 pig farmers in Hubei Province were selected as the research objects, and the Heckman two-stage model was used for empirical analysis, and the impact of health consciousness on different groups of large-scale pig farmers was discussed. The results showed that most scale farmers (78.97%) were willing to use environmental protection feed, and the average willingness to pay (WTP) was 6.69% (46.80 yuan each) of feed cost per pig. The level of health concern in health consciousness had a positive effect on scale breeding farmers to choose environmental protection feed, and the perception of health change had a positive effect on WTP of environmental protection feed. Through principal component analysis, it can be concluded that health consciousness had a direct and positive impact on the use and WTP for environmental protection feed on the whole. The difference analysis showed that there were significant differences among different groups of farmers with different health consciousness in the decision-making of willingness to pay for environmental protection feed. In view of this, this paper put forward some suggestions: government departments should strengthen environmental protection propaganda and promote the cultivation of health consciousness and environmental protection habits of large-scale farmers by various means; the characteristics of farmers as “rational economic man” should also be taken into account, and the adoption of environmental protection feed should become the norm of livestock industry by establishing and improving the corresponding reward and punishment system.

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Research progress of soybean combine harvester cleaning equipment and key technologies

LIU Peng, JIN Chengqian, YIN Xiang, NING Xinjie, LI Qinglun

2019, 31(10): 1758-1766. DOI: 10.3969/j.issn.1004-1524.2019.10.22

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Cleaning device is the main equipment of soybean combine harvester for cleaning and separating threshing mixture. Its performance determines the level of cleaning operation of soybean combine harvester. There are few cleaning devices specially used for mechanical harvest of soybean in China. At present, the combine harvester mainly adopts sieve-air cleaning device. This kind of cleaning device has a low matching with soybean characteristics, which leads to a high loss rate and impurity of soybean mechanical harvesting. In this paper, the research status of soybean combine harvester cleaning device was summarized from two aspects of cleaning device and key technologies, and the advanced technology of soybean combine harvester cleaning device in the world was summarized. The deficiencies of current research on soybean combine harvester cleaning device in China were analyzed, and the trend of future soybean combine harvester cleaning device was prospected.

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