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    20 May 2020, Volume 29 Issue 5
    Meta-analysis of the response to nitrogen deposition of soil nitrogen fractions, grassland soil C∶N∶P stoichiometry and microbial respiration
    YANG Nai-rui, HU Yu-fu, SHU Xiang-yang, ZENG Jian, ZHANG Xiang-lin, SHENTU Yu-cheng, HE Jia, CHENG Qi, LI Jie, LI Zhi, YU Ying
    2020, 29(5):  1-12.  DOI: 10.11686/cyxb2019374
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    This research studied the general response to nitrogen deposition, of soil N fractions, grassland C∶N∶P stoichiometry and microbial respiration. Data from 92 published articles from within China and abroad were analyzed by meta-analysis and the effects of nitrogen deposition rate, experimental duration and climatic conditions on nitrogen behavior were studied. It was found that, nitrogen deposition significantly increased soil total nitrogen (7.1%), available nitrogen (36.3%), ammonium nitrogen (51.3%), nitrate nitrogen (98.1%) and microbial biomass C∶P (53.6%), and significantly decreased soil pH (-5.9%), microbial biomass nitrogen (-11.5%), and microbial biomass phosphorus (-19.4%). With increase in nitrogen deposition rate, increases were seen in the size of the effect on total nitrogen (4.1%), soil N∶P (10.6%), dissolved organic nitrogen (41.8%), ammonium nitrogen (31.3%) and nitrate nitrogen (20.8%), while decreases were seen in soil pH (-5.8%), microbial biomass carbon (-13.2%), microbial biomass nitrogen (-11.5%), microbial biomass phosphorus (-18.0%), microbial biomass C∶P (-26.5%) and microbial respiration (-10.6%). As the experimental duration increased, the size of the nitrogen deposition effect on soil ammonium nitrogen (22.8%) significantly increased, while the effect on soil pH (-2.5%), microbial biomass carbon (-12.2%), microbial biomass nitrogen (-21.4%), and microbial respiration (-25.3%) significantly decreased. In addition, the mean annual temperature and precipitation significantly affected the response of microbial respiration to nitrogen deposition, but had no effect on the response of soil and microbial carbon, nitrogen and phosphorus to nitrogen deposition.
    Effects of the excavation activities of plateau zokor (Myospalax baileyi) on the plant community characteristics of alpine meadow
    YANG Zi-han, CHEN Tai-xiang, ZHENG Qiao-yan, WEI Wan-rong
    2020, 29(5):  13-20.  DOI: 10.11686/cyxb2019416
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    The excavation activities of plateau zokor can affect the grassland plant community composition and structure, resulting in spatial heterogeneity, namely the mounds and undisturbed vegetation patches. In this study, the vegetation characteristics and disturbance effects of different plateau zokor densities in winter pasture were evaluated, and the functional response of community composition and structure were assessed. With increased zokor population density, vegetation height increased significantly (P<0.05), the plant community cover and grass functional group species richness decreased (P<0.05), and the species richness and the forb richness initially increased and then decreased. Within increasing plateau zokor disturbance levels, the total underground biomass decreased (P<0.05), the biomass of the forbs increased (P<0.05), and the grass biomass initially decreased and then increased. These results indicate that moderate disturbance by plateau zokor enhances plant community species richness, while at low or high zokor density and disturbance, grass biomass is improved.
    Differences among three artificial grasslands in dynamics and community diversity of soil microarthropods
    QIU Yue, WU Peng-fei, WEI Xue
    2020, 29(5):  21-32.  DOI: 10.11686/cyxb2019444
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    Soil microarthropods are an important component of grassland ecosystems and can be used to monitor or evaluate the quality or health of grassland soils. The aim of this study was to evaluate the effects of different artificial perennial rangelands comprising plant species monocultures, on the diversity of soil microarthropods communities on the Qinghai-Tibetan Plateau. Three artificial perennial rangeland monocultures were established (Elymus sibiricus, Elymus nutans and Medicago sativa) in Hongyuan County, northwest Sichuan Province, and studied at three and five years from establishment, together with a natural grassland site (NG) for comparison. The investigations of the soil microarthropods communities and soil environment examined the 0-15 cm soil layer and were conducted in July and October 2018. Soil microarthropods were extracted from soil samples for 48 h at 38 ℃ in the laboratory, using the Tullgren. It was found that: 1) The community structure of soil microarthropods differed among the artificial grasslands and the main taxonomic groups affecting community structure were Tectocepheus, Petalomium and Hesperinidae. Microarthropod communities of three-year old artificial grasslands did not differ significantly (P>0.05) in density, group number and Shannon diversity index, while the density, group number and Shannon diversity index of microarthropod communities in NG and five-year old M. sativa rangeland were significantly (P<0.05) higher than those in five-year old E. sibiricus and E. nutans grasslands. In addition, the seasonal dynamics of soil microarthropod communities differed among the three artificial grasslands. 2) The density, group number and Shannon index of soil microarthropod communities decreased significantly in E. sibiricus and E. nutans grasslands (P<0.05) and increased in the M. sativa grassland (P>0.05) with increase in years established. 3) The main environmental factors affecting the composition of soil microarthropod communities were soil available P and C∶N and N∶P ratios. The soil available K significantly and positively correlated with the soil microarthropod density (P<0.05), and plant community coverage had a significant and positive effect on the group number and Shannon diversity index (P<0.05). The results show that the effects of the three evaluated artificial perennial rangeland monocultures on the community structure, density and diversity of soil microarthropods differ with forage species and establishment time. Appropriate measures should be taken to increase the soil microarthropod diversity of artificial E. sibiricus, E. nutans and M. sativa rangelands on the Qinghai-Tibetan Plateau where these are more than four years old, in order to promote normal ecological function.
    Effect of indoor-feeding at low altitude on muscle quality of yaks
    YANG Qin, GUAN Jiu-qiang, CHAI Zhi-xin, LI Hua-de, CAO Shi-xiao, ZHANG Xiang-fei, BAI Qin, ZHONG Jin-cheng, LUO Xiao-lin
    2020, 29(5):  33-42.  DOI: 10.11686/cyxb2019451
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    Indoor feeding of yaks at low altitude is an important means to overcome the winter forage supply problem for the large numbers of livestock in the Tibetan region, and it can also facilitate the growth and development of yak during the cold season and the improve the quality of yak meat. The purpose of our study is to explore the effects of indoor feeding in low-altitude areas on yak growth and development, slaughter performance, muscle quality, and serum indicators, and to provide preliminary data on factors that may influence yak muscle quality when indoor-feeding at low-altitude. Ten healthy 36-month-old male yaks of similar body weight and physical condition at 36 months of age were randomly divided into an indoor-feeding group and a grazing group. The indoor-feeding group were housed for fattening at Guanghan agricultural area (altitude: 600 m), and the grazing group were naturally grazed following traditional yak farming practice at 3500 m altitude in Hongyuan County. The experiment duration was 150 days. It was found that the weight gain of yaks fed indoors was significantly higher than that of the grazing group (P<0.01), and the yak carcass weight, pure meat weight, dressing-out percentage which means ratio of carcass weight to live weight and pure meat percentage were all significantly higher than that of the grazing group (P<0.01). The dietary quality of the longissimus muscle in the indoor-feeding group was significantly improved: The brightness (L*) was significantly higher than that in the grazing group (P<0.05), the cooking percentage which is also known as cooking loss and intramuscular fat content were significantly higher than those in the grazing group (P<0.05), and the shear force was significantly lower than the grazing group (P<0.05). The nutritional quality of the longissimus dorsi muscle in the indoor-feeding group was also significantly improved: The ether extract content was significantly higher than that in the grazing group (P<0.01). The contents of palmitic acid (C16:0) and stearic acid (C18:0) in the indoor-feeding group were extremely high compared to the grazing group (P<0.01), and the contents of saturated fatty acids, polyunsaturated fatty acids, and trans fatty acids were significantly higher than those in the grazing group (P<0.05). The flavor-forming free amino acid levels were significantly higher in the indoor-feeding group than in the grazing group (P<0.05). For serum indicators, the blood glucose concentration in the indoor-feeding group was significantly higher than that in the grazing group (P<0.05), while increase in triglyceride and total cholesterol levels, compared to the grazing group were highly significant (P<0.01). The concentration of β-hydroxybutyric acid was significantly lower than the grazing group (P<0.01); the acetyl CoA carboxylase enzyme activity was higher than the grazing group (P<0.01); the fatty acid synthetase enzyme activity was significantly higher than the grazing group (P<0.05); and the hormone-sensitive lipase (HSL) enzyme activity was significantly lower than the grazing group (P<0.05). The test results show that yak weight gain and fattening can be achieved by indoor feeding at low altitude in the cold season. Through activation of serum enzyme activity related to lipid deposition in yak, indoor-feeding can facilitate the synthesis and deposition of fat in muscle and improve the dietary quality and nutritional quality of yak meat.
    Nutritional value of forage sweet sorghum and its gas production performance evaluated using incubation with sheep rumen fluid in vitro
    ZHOU En-guang, WANG Hu-cheng, SHANG Zhan-huan
    2020, 29(5):  43-49.  DOI: 10.11686/cyxb2019354
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    This experiment studied the nutritional value and gas production performance, when incubated with sheep rumen fluid, of sweet sorghum forage harvested at different crop growth stages. The forage sweet sorghum cultivar Dajiang 3180 was harvested in August (S1), September (S2), October (S3) and November (S4), the herbage nutritive value analyzed. And the relative feeding value (RFV), total digestible nutrient (TDN), dry matter intake (DMI), digestible dry matter (DDM) and relative forage quality (RFQ) were evaluated. Herbage digestibility and the gas production at 3, 6, 9, 12, 24, 36 and 48 h were evaluated by in vitro simulated fermentation. It was found that: 1) There were significant differences in dry matter, crude protein, neutral detergent fiber, acid detergent fiber, ethanol extract and crude ash levels at the different harvest dates (P<0.05). 2) There were significant differences across the four harvest periods in RFV, DMI and RFQ (S1>S4>S3>S2) (P<0.001). The TDN and DDM of S1 and S4 were higher than the S2 and S3 harvests (P<0.001). 3) For the first 12 h of fermentation, there were gas production (GP) differences between the four harvest periods (P<0.001), with the GP of S3 being the highest. After 24 h, the GP of S4 was higher than S2 (P<0.05); There were no significant differences in GP between the four harvest periods after 36 h incubation (P>0.05). 4) There were no significant difference in DMD between the four harvest periods at 12, 24 and 48 h of incubation (P>0.05). In summary, the feeding value of sweet sorghum in the ‘hill and gully’ region of the Loess Plateau, China, is highest in August.
    Nutritional composition of forage rape and its rumen degradation characteristics in goats
    ZHAO Na, YANG Xue-hai, WEI Jin-tao, GUO Wan-zheng, CHEN Fang, ZHOU Guang-sheng, FU Ting-dong
    2020, 29(5):  50-57.  DOI: 10.11686/cyxb2019376
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    In this research, the nutritive value parameters of Huayouza 62 forage rape at flowering and early podding stages of crop development were evaluated by chemical analysis and rumen degradation characteristics were determined using three rumen-fistulated Boer goats. The results provide nutritional parameters for feeding forage rape to ruminants. It was found that: 1) The crude protein (CP) of forage rape in the flowering stage was 17.30%, and the total amino acid content was 11.48%, and both values were significantly higher than at the early podding stage (P<0.05). There were no significant (P>0.05) differences between the flowering and early podding crop growth stages in the contents of CP, crude ash (CA), calcium (Ca), total phosphate (TP), neutral detergent fiber (NDF), and acid detergent fiber (ADF). The glucosinolate content in the flowering stage was 10.17 μmol·g-1 and the tannin content was 0.94%, and these values were higher than those in early podding stage. 2) The rumen degradation rate of dry matter (DM), CP, organic matter (OM), NDF and ADF in the flowering and early podding stages gradually increased with increased digestion time. The CP rumen degradation rate of forage rape in flowering stage was 84.7% at 72 h. The DM rumen degradation rate increased rapidly from 0 to 24 h and stabilized at about 67% after 48 h, while the rumen degradation of NDF and ADF progressed more slowly, with values of 48.22% and 45.78%, respectively, at 72 h. The rumen degradation rate of DM and OM at the early podding stage was lower than at the flowering stage at the same time point from 0 to 72 h. 3) The rapidly soluble component of the various nutrient fractions differed significantly (P<0.05) between the flowering and early podding stage. CP was the nutrient fraction with the highest rapidly soluble values, reaching 64.06% at flowering and 68.81% at early podding, while the effective degradation rates of CP at these two growth stages were 76.50% and 75.13% (P<0.05), respectively. In conclusion, the feeding value of forage rape in both the flowering stage and the early podding stage is very high. The rumen degradation rate of the main nutrients is high, making forage rape suitable as a ruminant feed. Forage rape was more easily digested by goats in the flowering stage, than in the early podding stage.
    In vitro gas production characteristics and fermentation parameters of feedstuffs with varying proportions of cotton stalks and beet pulp
    LU Yan, ZHANG Ling-li, LUO Yuan-qin, WEI Li, XUE Xue, SUN Xin-wen, XIANG Chun-he, MAO Sheng-yong, WANG Xin-feng, ZHANG Wen-ju
    2020, 29(5):  58-66.  DOI: 10.11686/cyxb2019357
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    This experiment evaluated the nutritional quality, gas production characteristics and the fermentation products of mixed feedstuffs comprising cotton stalk and beet pulp. A three-factor, three-level orthogonal treatment combination and an in vitro fermentation method were used, in order to identify the optimal fermentation conditions. Beet pulp was added to cotton stalk in proportions of 10%, 30% or 50%, with 0.1%, 0.2% or 0.3% added urea, and 0.05%, 0.1% or 0.2% added bacterial liquid (lactic acid bacteria and yeast are the main components), with salt added 0.2% in all cases. From the 27 available combinations, 9 were chosen for inclusion in the experiment, designated A-I, and a control, K, as follows: A) 10% beet pulp, 90% cotton stalk, 0.1% urea, 0.05% bacterial liquid, 0.2% salt; B) 10% beet pulp, 90% cotton stalk, 0.2% urea, 0.1% bacterial liquid, 0.2% salt; C) 10% beet pulp, 90% cotton stalk, 0.3% urea, 0.2% bacterial liquid, 0.2% salt; D) 30% beet pulp, 70% cotton stalk, 0.3% urea, 0.05% bacterial liquid, 0.2% salt; E) 30% beet pulp, 70% cotton stalk, 0.1% urea, 0.1% bacterial liquid, 0.2% salt; F) 30% beet pulp, 70% cotton stalk, 0.2% urea, 0.2% bacterial liquid, 0.2% salt; G) 50% beet pulp, 50% cotton stalk, 0.2% urea, 0.05% bacterial liquid, 0.2% salt; H) 50% beet pulp, 50% cotton stalk, 0.3% urea, 0.1% bacterial liquid, 0.2% salt; I) 50% beet pulp, 50% cotton stalk, 0.1% urea, 0.2% bacterial liquid, 0.2% salt and control group K (100% cotton stalk, 0 urea, 0 bacterial liquid, 0.2% salt). It was found that for gas production at 48 hours, Group I was significantly higher than groups A, D, F (P<0.05), and group K (P<0.01), while, there were no significant difference between the other groups (P>0.05). Group I had the highest organic matter digestibility and metabolizable energy: 722.40 g·kg-1 and 9.75 MJ·kg-1, respectively. The slow degradation parameter, b, was significantly higher in group I than in groups A, D, F (P<0.05), and group K (P<0.01). Groups G and H, were also significantly higher than group K (P<0.05), while there were no significant differences between the other groups (P>0.05). The rapid degradation parameter (a), and the gas production rate (c), did not differ significantly between any of the groups. The concentration of ammonia nitrogen in group H was significantly lower (P<0.05) than in other groups, and group K was significantly lower than groups A, B, C and E. Soluble sugar was significantly higher (P<0.05) in groups H and I than other groups, and in group H was 44.06% and 31.23% higher than the lowest groups, B and K, respectively. No significant differences between groups for pH, lactic acid and microbial protein were detected (P>0.05). The highest total volatile fatty acid (VFA) concentrations occurred in groups was group H and G (100.31 and 99.73 mmol·L-1, respectively), and these values were which was significantly (P<0.05) higher than those of other groups. For the volatile fatty acids, acetate and butyrate were highest in Groups G, H, and I and propionate was highest in Groups G and H, while acetate:propionate ratio was lowest in groups G, H, and I. Using a membership function analysis for multivariate ranking of the mixed fermentation products of cotton stalk and beet pulp, the average membership function values of each group were I (0.719)>H (0.692)>F (0.595)>G (0.591)>C (0.407)>E (0.400)>B (0.395)>K (0.374)>D (0.307)>A (0.243). In conclusion, a dry matter-based ratio of cotton stalk to beet pulp of 50∶50, with 0.1% urea, 0.2% bacterial liquid, and 0.2% salt (group I) is recommended based on this research. The mixed fermentation product of this mixture has the highest in vitro gas production, the highest digestible organic matter and metabolic energy content, the higher concentration of acetic acid and butyric acid, and the highest multivariate score of the fermentation products. It is the best among the fermentation combinations tested and is suitable as a roughage for ruminants.
    Analysis of codon usage bias in the genome of Epichloё gansuensis
    LI Xiu-zhang, SONG Hui, ZHANG Zong-hao, XU Hai-feng, LIU Xin, LI Yu-ling, LI Chun-jie
    2020, 29(5):  67-77.  DOI: 10.11686/cyxb2019335
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    The sequence of multiple sets of triplet adjacent DNA or RNA nucleotides (codon) determines amino acid sequence. In the long process of adaptive evolution, the codon usage of organisms has developed certain biases. Two major factors affect codon usage bias. The species ‘Epichloё gansuensis’ is an important endophytic fungus. E. gansuensis infection can improve the host resistance to biotic and abiotic stress, but the fungus produces secondary alkaloid metabolites that may poison livestock. This study aimed to analyze the basic composition of the genome of E. gansuensis and the pattern of codon usage, and to determine the optimal codon. Correlation of each codon index with gene length, amino acid usage frequency and other indicators was analyzed, to identify the main factors affecting codon usage. We found that the GC3 is mainly distributed between 40%-60%, the effective codon number of 62.7% genes distributed between 0-0.1, 95.4% effective codon numbers are lower than the expected and lie to the lower right of the expectation curve. These results suggest that mutation pressure is one of the main influencing factors on codon usage of E. gansuensis. However, the neutral analysis showed a significant positive correlation between GC12 and GC3 (r=0.110, P<0.01), with a slope of the regression line 0.2729. It suggests that the effect of mutation on codon usage was only 27.29%, 82.71% for other factors. Correlation analysis also showed that hydrophobic amino acids had the greatest effect on the codon usage of E. gansuensis, greater than that of aromatic amino acids. In addition, 26 optimal codon that end with a G or C residue from E. gansuensis were identified, indicating that codon usage in E. gansuensis may relate to the presence of a GC residue at the third position of the triplet codon. These optimal codon are significantly correlated with transcription level, and may help in the design of degenerative primers, for tracing the evolutionary origin of endophytic fungi.
    Arbuscular mycorrhizal fungal infection rates of flora of the Fengfeng mining area coal gob piles and influence on plant Cd content
    ZHAO Xin, WU Zi-long, ZHANG Hao, YANG Xu-zhao, HAN Chao, GAO Jie
    2020, 29(5):  78-87.  DOI: 10.11686/cyxb2019528
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    This research studied arbuscular mycorrhizal fungus infection rates in plants and soil collected from the coal gob piles at the Fifth, Jiulong and Wutongzhuang mines in the Fengfeng mining area, and the Cd levels measured using graphite furnace atomic absorption spectrophotometry. The Cd enrichment ability was also compared by calculating transport and enrichment coefficients. There were 20 plant species belonging to 19 genera and 7 families around the coal gob piles at the three mines. The plant families present were: Gramineae, Asteraceae, Moraceae, Asclepiadaceae, Chenopodiaceae, Polygonaceae and Amaranthaceae. Setaria viridis was the most commonly occurring species. All plants at the three mines were infected by arbuscular mycorrhizal fungi, and formed mycorrhizal symbioses but the fungal mycelium and vesicle densities were low for the majority of plants. The species with the highest average infection rate for the three mines was S. viridis (59.22%) and the second highest was Metaplexis japonica. The Soil Cd concentrations at the three mines were similar, ranging from 1.778 to 2.066 mg·kg-1 on average. Plant Cd content differed between aboveground and underground plant organs. For aboveground herbage of plants from the Fifth and Wutongzhuang mines, the highest Cd content was observed in Artemisia scoparia (0.863 and 0.536 mg·kg-1, respectively) and the lowest Cd content was in Chloris virgata and Portulaca oleracea (0.253 and 0.203 mg·kg-1, respectively, for the two mines). Therefore the highest plant Cd concentrations were 3.41 times and 2.64 times the lowest, respectively, at the two mines. The plant aboveground herbage Cd concentration at the Jiulong mine was highest in S. viridis (0.913 mg·kg-1), and this was significantly higher than that of other plants, and was 7.94 times higher than herbage Cd concentration in Cynodon dactylon which had the lowest Cd content. The content of Cd in the underground organs of plants from three mines was the highest in S. viridis. The belowground to aboveground transfer coefficients of the plants from the three mines were higher than their own enrichment coefficients. The transfer coefficient of some plants reached or even exceeded 1, but the enrichment coefficients were all less than 1. For the three mines, the transfer coefficients of A. scoparia and Fagopyrum tataricum were highest, while the enrichment coefficients of A. scoparia and S. viridis were higher than those of other plants. A correlation analysis showed that arbuscular mycorrhizal fungal infection promoted the absorption and enrichment of Cd from the soil by the roots of S. viridis. Arbuscular mycorrhizal fungi were therefore beneficial to the remediation of heavy metal contaminated soil. This study thus provides reference data for repair of soil heavy metal pollution coal gob piles in the Fengfeng coal mining area, using locally dominant plant species and arbuscular mycorrhizal fungi to enhance Cd absorption.
    Isolation, identification and biological characteristics of Alternaria brassicicola leaf spot on Orychophragums violaceus
    WANG Chun-ming, YUAN Wei-wei, ZHANG Xiao-jie, ZHOU Tian-wang, GUO Cheng, JIN She-lin
    2020, 29(5):  88-97.  DOI: 10.11686/cyxb2019362
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    A leaf spot disease was found on Orychophragums violaceus in Huanghua City, Hebei Province, during October of 2017. The infected leaves were initially black and brown with irregular small disease spots, which gradually developed into necrotic spots with concentric circles, affecting 96.2% of plants observed and 87.6% of leaves on each plant. In order to identify the pathogen, samples were isolated from infected leaves by tissue separation and cultured. After purification, three isolates were obtained (GS1-1, HH2-1, and HH3-1), and their pathogenicity tested according to Koch’s rule. Test results confirmed that the three isolates reproduced the original symptoms and were therefore the causative agent of the observed O. violaceus leaf spot disease. For the three isolates, DNA was extracted, amplified and sequenced by rDNA-ITS, and the results showed the three isolates had a very close relationship to Alternaria brassicicola (MN173824, MN173825, MN173823 and MF462311), and 100% similarity with A. brassicicola (MF462311). The results also showed a very close relationship of the three isolates to certain other A. brassicicola isolates (JX213350, KF889266, KT895946, KC584642, LC480212 and KT895946) through EF-1α gene sequencing, which was consistent with the results of morphological identification. The vegetative growth and sporulation conditions of isolate GS1-1 were studied by biological methods. It was found that the optimal medium for vegetative growth and spore production of isolated GS1-1 was PSA, and the optimal carbon and nitrogen sources for vegetative growth were maltose and peptone, respectively. The optimal carbon and nitrogen sources for sporulation were glucose and yeast extract, respectively. The optimal colony growth temperature was 25 ℃, with the temperature range over which colony growth occurred being 5-35 ℃, and the optimal spore production temperature was 28 ℃. Alternate 12 h light and dark periods promoted colony growth and sporulation.
    Effect of water and fertilizer coupling on Leymus chinensis in the Ningxia irrigation area
    FU Bing-zhe, ZHOU Yan-fei, LI Xue, NI Biao, GAO Xue-qin
    2020, 29(5):  98-108.  DOI: 10.11686/cyxb2019364
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    This research investigated the effects of water and fertilizer and their interaction on the yield, hay quality and seed yield of Leymus chinensis in the Ningxia irrigation area. The L. chinensis cultivar used was Zhongke No.2. A two factor split-plot experiment design was adopted, with water as the main plot treatment and fertilizer as the sub-plot treatment. A quadratic polynomial stepwise regression and normalization was used to identify the combination of irrigation and fertilizer that best optimized the multi-factor benefit. It was found that irrigation had significant effects on seed and hay yield, fertilizer partial productivity (PFP), irrigation water use efficiency (iWUE), and hay quality (P<0.05). Fertilization had significant effects on seed and hay yield, PFP, quality, and hay iWUE (P<0.05); significant (P<0.05) water fertilizer interactions were observed for thousand-grain weight, heading rate, hay quality, hay PFP and iWUE. The seed yield was highest with 360 mm irrigation and 132 kg·ha-1 fertilizer application. The hay yield and relative feeding value (RFV) were highest with 360 mm irrigation and 540 kg·ha-1 fertilizer application. Stepwise analysis showed that the interaction effect of water and fertilizer differs depending on the purpose of L. chinensis cultivation. When L. chinensis is planted for hay production and forage quality is important, the recommendation is to apply 288-360 mm and 324-540 kg·ha-1 of fertilizer; when L. chinensis is planted for seed production and when efficient use of fertilizer is important, the recommendation is to apply 288-360 mm irrigation and 108-216 kg·ha-1 of fertilizer.
    Effects of different rhizobium fertilizers on alfalfa productivity and soil fertility
    HE Guo-xing, SONG Jian-chao, WEN Ya-jie, LIU Cai-ting, QI Juan
    2020, 29(5):  109-120.  DOI: 10.11686/cyxb2019344
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    After potting and field experiments to screen for the best match with Medicago sativa cv. Gannong No.3 and Gannong No.9, six rhizobium strains (Da99, DL58, G3G2, Ga66, QL31B and Wa32) were selected for further research and prepared as solid rhizobium fertilizers. In April 2017, the rhizobium fertilizers were tested on a second-year stand of M. sativa cv. Gannong No.5, to study their effect on plant productivity and soil fertility in an alfalfa crop. Non-fertilizer was used as control. It was found that strain Da99 produced the greatest effects on plant dry weight and height (increases of 41.75% and 9.30%, respectively; P<0.05), compared with the control. However, none of the rhizobium fertilizers significantly affected the stem∶leaf ratio. The strains QL31B, DL58 and G3G2, significantly lowered water content of the air-dried soil, compared to the control. Differences rhizobium fertilizer reduced the soil pH. Treatment with Wa32 increased soil organic matter, alkaline nitrogen and available phosphorus contents by 75.39%, 22.79% and 76.63%, respectively, compared to the control. Treatment with Da99 and G3G2 strains significantly increased soil available potassium levels by 27.35% and 24.03%, respectively, compared to the control. Some rhizobium fertilizer treatments improved soil urease activity, but effects on soil catalase activity were not significant. Treatment with QL31B increased the soil sucrase and alkaline phosphatase activities by 76.94% and 21.81%, respectively, compared to the control. The soil cellulase activity after treatment of G3G2 and Ga66 was increased significantly by 22.35% and 18.82%, respectively, compared to the control. Multivariate evaluation showed that treatment with the six rhizobium fertilizers significantly improved the productivity and soil fertility of alfalfa. The best performing rhizobial strain, with the most obvious effect on plant growth and soil fertility in the current study, was QL31B, which is therefore preliminarily concluded to be a broad-spectrum rhizobium.
    Effects of the combined application of biochar and chemical fertilizer on fertility and microbial characteristics of purple soil and yield and quality of oilseed rape
    XU Qi-wen, MA Shu-min, ZHU Bo, ZHANG Xiao-duan, XING Yi, DUAN Mei-chun, WANG Long-chang
    2020, 29(5):  121-131.  DOI: 10.11686/cyxb2019338
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    The aim of this study was to investigate the effects of the combined application of biochar and chemical fertilizer on fertility and microbial characteristics of ‘purple soil’ (an Entisol in USDA taxonomy) and yield and quality of oilseed rape, so as to provide a theoretical basis for formulating an optimal combination of biochar and chemical fertilizer in Southwest China. The treatments included: B0F1 (no biochar+reduced fertilization), B0F2 (no biochar+traditional fertilization), B1F1 (low biochar+reduced fertilization), B1F2 (low biochar+traditional fertilization), B2F1 (high biochar+reduced fertilization), and B2F2 (high biochar+traditional fertilization), arranged in a split-plot design. It was found that soil pH, cation exchange capacity (CEC), total nitrogen (TN), total carbon (TC) and total potassium (TK) increased significantly under B1 and B2, compared with values for B0, while no significant difference was detected between F1 and F2. Biochar application promoted diversity of soil microorganisms with increased Shannon’s index (H), Simpson index (D) and abundance index of carbon source utilization (S), and improved the overall utilization ability of carbohydrates, amino acids, carboxylic acids, polymers and phenolic compounds. Yield of oilseed increased significantly with application of biochar. Contents of erucic acid, glucosinolates and linolenic acid decreased, while levels of oleic acid and yellowness increased with application of biochar under F1. Redundancy analysis showed that factors influencing oilseed quality included, in order of significance, TK, TC, TN, total phosphorus (TP), CEC and soil pH, among which TK, TC, TN and TP affected oilseed quality significantly (P<0.05). In conclusion, the combined application of biochar and chemical fertilizer significantly improved the fertility status of soil, increased the activity of soil microorganisms, and promoted the microbial utilization of carbon sources. The optimal application of biochar was 35 t·ha-1. Biochar at this rate effectively offset the nutrient deficiency after the reduction of fertilizer, and improved the yield and quality of oilseed rape.
    Effects of different direct seeding methods and weed control timing on weed occurrence and grain yield of early indica rice
    ZHOU Yan-zhi, CHEN Li-ming, WANG Wen-xia, LI Zu-jun, ZENG Yong-jun, TAN Xue-ming, HU Shui-xiu, SHI Qing-hua, PAN Xiao-hua, ZENG Yan-hua
    2020, 29(5):  132-140.  DOI: 10.11686/cyxb2019421
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    This research aimed to identify the optimal timing of weed control in rice paddy fields. Hybrid early indica rice Qiliangyou 2012 was planted in pot experiments with three sowing methods [dry direct seeding (DDS), wet direct seeding (WDS) and flooding direct seeding (FDS)] and four manual weeding treatments [from sowing to three leaves (C1), from three leaves to four leaves (C2), weeding for the whole growth period (C3) and no weeding over the entire growth period (CK)], to study the effects of the different seeding methods and weed control options on weed occurrence and grain yield. It was found that in the DDS treatment, the seeding emergence rate was significantly increased compared to FDS and WDS. Weeds such as Echinochloa crusgalli and Euphorbia lathyris were a serious problem in DDS, while aquatic weeds including Juncellus serotinus and Monochoria vaginalis were relatively serious in FDS and WDS. The weed occurrence in the DDS treatment was serious in early crop development (3-29 days after sowing), but was significantly lower than that of FDS and WDS after 43 days. For the whole rice growing period, the weed density in DDS was decreased by 12.57%-15.72%, and the biomass of weeds was significantly lower than that of FDS and WDS, while the fresh weight of weeds in DDS was decreased by 65.83% and 57.45%, while the dry weight was decreased by 69.73% and 59.04%, respectively, compared to FDS and WDS. Correlation analysis showed that weed biomass was negatively correlated with grain yield and its components for direct seeding rice, and the occurrence of weeds 15-22 days after sowing significantly reduced yield. Compared with CK, the grain yield of early indica rice was significantly increased by all weed control treatments, with the ranking for yield being C3>C1>C2>CK, and the average yield increase being 17.91%-31.21%. The best among the weed control treatments was C1; while the yield of DDS was higher than the yields of FDS and WDS, and the increase in grain yield was mainly due to an increase in number of effective panicles per unit area. The peak occurrence of weeds occurred at different times in the different sowing treatments. DDS has potential to reduce the occurrence of weeds in direct seeded paddy fields, and used with weed control from sowing to the three leaf stage, could enhance rice crop yield.
    Effects of planting density and nitrogen fertilizer rate on maize yield and photosynthetic characteristics in arid areas of central Gansu, China
    KANG Cai-rui, XIE Jun-hong, LI Ling-ling, WANG Jia-nan, GUO Xi-jun, PENG Zheng-kai, WANG Jin-bin, Setor kwami Fudjoe, WANG Lin-lin
    2020, 29(5):  141-149.  DOI: 10.11686/cyxb2019342
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    This research evaluated the effects of planting density and nitrogen fertilizer rate on maize photosynthetic characteristics, yield and yield components under full film double ridge plastic mulch. The research relied on a field experiment that was set up in 2012 at Dingxi, in the Longzhong region. The experiment used a split plot design, with four planting densities as the main plot factor: 45000 (D1), 52500 (D2), 60000 (D3), 67500 plants·ha-1 (D4). The sub plot treatment comprised two nitrogen fertilizer rates: 200 (N2) and 300 kg N·ha-1 (N3). It was found that the leaf area index of maize improved with increase in planting density and nitrogen fertilizer rate, while the chlorophyll concentration decreased with increasing planting density. With increased planting density, the photosynthesis rate of maize decreased. Nitrogen fertilizer rates had no significant effect on the photosynthesis of maize. The treatment (D2N2) had the highest yield. The grain yield and biomass of D2 were, respectively, 15.2% and 14.5% higher than D1, while the grain yield and biomass under N2 were, respectively, 10.9% and 4.8% higher than those of N3. There was a significant positive correlation between grain yield and ear number and kernel number, but no significant correlation with hundred-grain weight. Therefore, under double furrow sowing with full film mulch in dryland farming areas of central Gansu, China, a planting density of 52500 plants·ha-1 and nitrogen fertilizer application of 200 kg N·ha-1 was found to be optimal for sustainable cultivation of maize. This cultivation regime facilitated leaf photosynthesis, leading to increased numbers of ears and kernels, thus enhancing maize grain yield.
    Growth, osmotic adjustment and antioxidant capacity responses of Schizonepeta tenuifolia to drought stress
    LI Ke, ZHOU Zhuang-yu, LI Si-ju, YAO Hao-zheng, ZHOU Ying, MIAO Yu-jing, TANG Xiao-qing, WANG Kang-cai
    2020, 29(5):  150-158.  DOI: 10.11686/cyxb2019368
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    This research studied the changes (relative to unstressed plants) in the growth, osmotic adjustment and antioxidant system of Schizonepeta tenuifolia grown in pots under severe or moderate water deficit stress [substrate relative water content 5%-10% (severe stress), 45%-50% (moderate stress) or 85%-90% (no stress, CK)], imposed for a duration of 15, 30 or 45 days. The growth of S. tenuifolia showed increasing inhibition with increasing intensity of water deficit, with traits affected including the growth rate, leaf dry weight and total chlorophyll content, while leaf mass per area (LMA) increased under water deficit. Water deficit promoted the accumulation of osmotic adjustment substances in leaves, including soluble sugar, soluble protein, proline and Na+. Water deficit also generally increased the antioxidant capacity of leaves: The activities of superoxide dismutase (SOD) and catalase (CAT) were moderately increased at 15 and 30 days, but decreased at 45 days, relative to CK, while peroxidase (POD) activity was moderately increased at 30 and 45 days. The contents of total phenol and total flavonoids were moderately increased at 15 days, and reached their highest levels under moderate drought at 30 and 45 days. Increased duration of water deficit affected the osmotic adjustment in leaves: The soluble sugar and soluble protein contents increased; the proline, K+, Na+ and Ca2+ contents decreased; the activities of SOD and CAT increased initially and later decreased; POD activity continued to increase with time, and the accumulation of total phenols and total flavonoids increased initially and then remained unchanged. In summary, S. tenuifolia made adaptive responses to drought stress and showed tolerance to moderate drought. This study provides a theoretical basis for the standardized cultivation and water management of S. tenuifolia.
    Cloning and functional analysis of the gene NtUFGT in Nitraria tangutorum
    GAO Zi-qi, WANG Jia, TANG Yu-chen, WANG Ying-chun
    2020, 29(5):  159-170.  DOI: 10.11686/cyxb2019334
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    Anthocyanidins are important secondary metabolites of flavonoids. Their significant antioxidant effects can assist plants to alleviate oxidative damage caused by various abiotic stressors. Based on transcriptome data, a flavonoid 3-O-glucosyltransferase gene was cloned from the cDNA of Nitraria tangutorum, and was named NtUFGT. The open reading frame of NtUFGT has 1407 bp, and encodes a protein with 468 amino acids. The relative molecular mass of the protein encoded by this gene is predicted to be 51.37 kDa. Multiple sequence alignment analysis showed that the NtUFGT protein belongs to the UDP-glycosyltransferase protein family. Quantitative real-time PCR (qRT-PCR) was carried out to analyze gene expression patterns. NtUFGT had tissue-specific expression patterns. From high to low expression: flower>fruit>stem>leaf>root. Also, it was rapidly up-regulated by polyethylene glycol and abscisic acid treatments. The plasmid of pPZP221:35S:NtUFGT was constructed and introduced into a wild-type (WT) Arabidopsis thaliana by a floral-dip method. qRT-PCR was carried out to confirm that the NtUFGT gene was expressed in three T3 generations. The growth status and stress-resistant physiological and biochemical indexes of wild-type and transgenic A. thaliana (OE lines) under drought stress were measured. The OE lines showed a stronger phenotype with longer root length, heavier fresh weight and higher chlorophyll content than the WT. OE lines also accumulated more flavonoids and anthocyanin. Consistent with phenotype, OE lines had increased levels of antioxidant enzyme (superoxide dismutase, peroxidase, and catalase) activity, and accumulated more glutathione, proline and less malondialdehyde and H2O2. The results of qRT-PCR analysis showed that the expression levels of the stress-related genes (AtCAT1, AtPOD1, AtRD29A) and the proline synthesis gene AtP5CS in OE lines were significantly higher than those in the WT. Our observations indicate that the NtUFGT can facilitate stronger active oxygen scavenging capacity and osmotic adjustment ability by effective promotion of the synthesis of anthocyanidins and flavonoids in transgenic Arabidopsis, thus enhancing the tolerance of plants to drought stress.
    Cloning and functional analysis of chlorophyllide a oxygenase encoding gene MtCAO in Medicago truncatula
    LIU Wen-wen, CUI Hui-ting, WEI Chun-xue, LONG Rui-cai, KANG Jun-mei, YANG Qing-chuan, WANG Zhen
    2020, 29(5):  171-181.  DOI: 10.11686/cyxb2019447
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    Plant biomass is determined by photosynthesis efficiency, which in turn is affected directly by the chlorophyll concentration. Chlorophyllide a oxygenase (CAO) is one of the three speed-limiting enzymes in the chlorophyll synthesis pathway, and catalyzes chlorophyllide a into chlorophyllide b. In this study, the open reading frame of Medicago truncatula CAO (MtCAO), which encodes a protein of 531 amino acid residues, was cloned. Phylogenetic analysis demonstrated that MtCAO was closer to its dicot homologs than to those in more distantly related plant species. Different from CAOs in lower plants, CAOs in higher plants contain nine exons. MtCAO was preferentially expressed in green tissues, especially leaves. When transiently expressed, the MtCAO-GFP recombinant protein was localized in the chloroplast of Arabidopsis protoplast cells. The transcript of MtCAO was repressed by the treatment with methyl jasmonate (100 μmol·L-1) or darkness, but induced by NaCl (100 mmol·L-1) or PEG (5%) within 4 h then repressed until 12 h from application. Also, the expression of MtCAO fluctuated with the circadian clock. These results are consistent with the cis-element analysis of the MtCAO promoter. MtCAO was ecotopically expressed in Arabidopsis. The analysis of two independent transgenic lines revealed no significant difference in the content of chlorophyll, the chlorophyll a∶b ratio or shoot weight from the wild type. The results are similar to those from AtCAO overexpression Arabidopsis. Therefore, we postulate that the non-abnormal phenotype of the transgenic Arabidopsis constitutively expressing MtCAO probably results from the fact that the conserved A domain contains a “degron” motif, which was proven to hamper the elevation of AtCAO in Arabidopsis overexpressing AtCAO.
    A high-throughput sequencing evaluation of bacterial diversity and community structure of the desert soil in the Junggar Basin
    WEI Peng, AN Sha-zhou, DONG Yi-qiang, SUN Zong-jiu, Bieerdawulieti·Xihayi, LI Chao
    2020, 29(5):  182-190.  DOI: 10.11686/cyxb2019337
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    Desert grassland accounts for 46.9% of the total grassland area in Xinjiang, with different desert types in areas of different climate, each with significantly different vegetation and soil types. Three representative desert vegetation types: Seriphidium transiliense desert in the southern margin, Haloxylon persicum desert in the hinterland and Anabasis salsa desert in the northern margin of the Junggar Basin, were chosen for study. The bacterial community composition and diversity characteristics of the three desert vegetation types were analyzed and compared using a high-throughput sequencing technique. It was found that Actinobacteria, Proteobacteria, Chloroflexi, Acidobacteria, Gemmatimonadetes, and Bacteroidetes were the six dominant groups of bacteria in these Junggar Basin soils. The cumulative abundance was more than 94%. Actinobacteria had the highest relative abundance (>50%) in all three desert types, and the difference between desert types for relative abundance of Actinobacteria was not significant (P>0.05). Acidobacteria had a relative abundance in S. transiliense desert notably higher than in other desert types, while Firmicutes bundance was higher in H. persicum desert than in other desert types, and Bacteroidetes had the highest relative abundance in A. salsa desert, and these differences were significant (P<0.05). Among the three desert types, the number of bacterial operational taxonomic units in H. persicum desert was the lowest and the community diversity was the lowest. Pearson correlation and redundancy analyses showed that average annual precipitation and soil organic carbon are the most significant environmental factors affecting the composition and diversity of the bacterial communities studied.
    Dynamics of disease and sporangium quantity, key factors analysis of grape downy mildew initial occurrence in Tianshui area
    DU Hui, JIANG Jing-jing, WANG Chun-ming, GUO Jian-guo, QI Yong-hong, LV He-ping, CHEN Ming
    2020, 29(5):  191-197.  DOI: 10.11686/cyxb2019372
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    Grape downy mildew is a major disease on grapes in Tianshui area. To clear the temporal dynamics of sporangium quantity and disease, the key factors to the disease initial occurrence, sporangia of Plasmopara viticola dispersal were observed using the spore-trapping machine and the disease state were investigated during the grape growing season in Tianshui area. At the same time, meteorological data were recorded in the field by automatic microclimate recording instrument. The results showed that the sporangium of P. viticola in Tianshui area was generally observed around the beginning of July. If conditions were suitable, it would be the quickly spreading period from late July to late August, and would enter the rapid decreasing period after September. The disease in the field initially occurred in the early or middle July, and reached its peak in the middle or late August when the conditions were suitable. Disease began to appear about 7 d after the sporangia of P. viticola were detected in the field, and there was a significant positive correlation between the disease index and the number of sporangium. The grape downy mildew is closely related to the meteorological factors of temperature, relative humidity, and rainfall, effective rainfall is the key factor to disease initial occurrence and spreading.