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    20 December 2015, Volume 24 Issue 12
    Orignal Article
    Soil respiration dynamics and impact factor analysis of a shrubland-grassland ecotone in the Eastern Qilian Mountains
    WEI Wei, ZHOU Juan-Juan, CAO Wen-Xia, XU Chang-Lin
    2015, 24(12):  1-9.  DOI: 10.11686/cyxb2015019
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    This research explored the factors influencing soil respiration dynamics of an alpine Shrubland-Grassland ecotone. The soil respiration dynamics were measured using a LI-8100A automated soil CO2 flux system, and relationship between soil respiration and soil factors was analysed for an eastern Qilian Mountain typical Shrubland-Grassland ecotone. The mean soil respiration rate was 2.3-7.2 μmol CO2/(m2·s). The rankings of soil respiration rate at different sampling points were: Polygonum viviparum meadow center (S1)>Polygonum viviparum meadow-Potentilla fruticosa shrub ecotone (MSC1)>Potentilla fruticosa shrub center (S2)>Potentilla fruticosa-Rhododendron shrub ecotone (MSC2)>Rhododendron shrub center (S3). The diurnal variation of soil respiration of S1 and MSC1 had a unimodal pattern, with the peak at 14:00. For S2, MSC2 and S3 respiration rates were only 0.3-1.1 μmol CO2/(m2·s) and the peak was not obvious. Soil physical properties showed a significant vertical gradation within ecotones. Soil respiration rate was significantly and positively correlated with soil temperature and total phosphorus (P<0.01), significantly negatively correlated with soil moisture (P<0.01), and significantly positively correlated with soil organic carbon in the 0-20 cm soil layer (P<0.05). Analysis of the relationship between soil respiration and soil temperature, soil moisture, total phosphorus, and organic carbon showed that soil temperature, soil moisture and total phosphorus were the main limiting factors for soil respiration rate. The highest fitted coefficient was soil temperature, which explained 79.9% of the spatial variability in soil respiration.
    Relationship between species diversity, productivity, climatic factors and soil nutrients in the desert steppe
    SUN Xiao-Li, KANG Sarula, ZHANG Qing, CHANG Chang-Ming, MA Wen-Jing, NIU Jian-Ming
    2015, 24(12):  10-19.  DOI: 10.11686/cyxb2015014
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    The effect of species diversity on ecosystem function is one of the core issues in ecology. Our objective was to explore the relationship between species diversity, productivity, climatic factors, and soil nutrients in the desert steppe, to enhance understanding of the diversity-productivity relationship. Species richness and Shannon-Wiener index were used to measure species diversity. Climatic, vegetation and soil data were collected for 80 sampling sites across the desert steppe of Inner Mongolia. Regression analysis and stepwise regression analysis were used for determining the relationship between environmental factors and species diversity and productivity. The effects of environmental factors on species diversity and productivity were evaluated, based on the coefficient of determination from regression analysis. It was found that species diversity and biomass were both positively and linearly correlated with mean annual precipitation and aridity index. However, Shannon-Wiener index and biomass were both negatively and linearly correlated with mean annual temperature. Species richness was negatively and linearly correlated with soil total phosphorus content (R2=0.072, P<0.05), but positively and linearly correlated with soil available nitrogen content (R2=0.183, P<0.01); Shannon-Wiener index was also positively and linearly correlated with soil available nitrogen content (R2=0.061, P<0.05), and biomass was positively and linearly correlated with total nitrogen, soil available nitrogen, and soil organic carbon content. The coefficients of determination were 0.150, 0.177, 0.089 (P<0.01), respectively. Biomass increased significantly with the increase of species richness and Shannon-Wiener index. These results indicate that aridity index is the most influential factor affecting species diversity and biomass of desert steppe.
    Effects of nitrogen and phosphorus on the soil nematode community in Tibetan Plateau alpine meadows
    WANG Jing, HU Jing, DU Guo-Zhen
    2015, 24(12):  20-28.  DOI: 10.11686/cyxb2015035
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    Soil nematodes are sensitive to environmental changes and are widely used as indicators of soil conditions. Here we monitored temporal dynamics of the soil nematode community using a multi-level N and P addition experiment in the eastern Tibetan Plateau alpine meadow under grazing conditions. The results showed N and P fertilizer applications slightly improved the diversity (H') and the total number of soil nematodes, but that the maturity index (MI) declined with increases in the fertilizer gradient. The abundance of fungivores and predators decreased whereas bacterivorous nematodes significantly increased after P input. With lower-level applications of N and P fertilizer, the abundance of herbivorous nematodes was low but that of omnivores was high. The abundance of cp-1 (colonizer-persister) and cp-3 group nematodes increased with increases in the fertilizer gradient. The largest abundance of cp-1 and cp-3 appeared in treatments with moderate N and P fertilizer applications. We found a significantly positive relationship between bacterivorous nematodes, cp-1 and available phosphorus. The response of soil nematodes to different kinds of fertilizer is probably caused by grazing.
    Soil moisture and infiltration characteristics for artificial pasture planted on opencast coal mining tailings
    YANG Zheng, WANG Dong, LIU Yu, ZHU Yuan-Jun, WU Gao-Lin
    2015, 24(12):  29-37.  DOI: 10.11686/cyxb2015231
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    This study analyzed the above-ground biomass, soil moisture and infiltration rates for different artificial pastures planted on opencast coal mine tailings. The above-ground biomass of different artificial grasslands was significantly different (P<0.05), with the above-ground biomass of shrub-grassland and mixed planting of shrub and grass being greater than that of single-plant pasture. The above-ground biomasses of Agropyron mongolicum+Artemisia desertorum pasture and Astragalus adsurgens+A. desertorum pasture were higher than those of A. mongolicum and A. adsurgens pasture by 40.84%-47.88% and 27.31%-53.49%, respectively. Soil water content of artificial pasture increased with increased soil depth and was highest for the 20-30 cm soil layer in Hedysarum scoparium pasture. The above-ground biomass of different artificial grassland types negatively correlated with the accumulated mean value of soil moisture. The mean initial infiltration of mixed plantings of shrub and grass were higher than those of single plant pasture by 44.25%. These results indicate that mixed planting of shrub and grass is the best choice for soil development and vegetation restoration of opencast coal mine tailings.
    Effects of different crop-mulberry intercropping systems on nutrients in arid purple soils in the Three Gorges Reservoir Area
    LIU Yue-Jiao, NI Jiu-Pai, ZHANG Yang, ZHOU Chuan
    2015, 24(12):  38-45.  DOI: 10.11686/cyxb2015008
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    To help address the problem of purple soil erosion on sloping land in Chongqing city, an experiment has been undertaken to investigate the effects of different crop-mulberry intercropping systems on soil nutrients. Four treatments were designed: crops were planted along transverse slopes (CT), crops were planted along transverse slopes with mulberry trees on the upper and lower slopes (T1), crops were planted along transverse slopes with mulberry trees on the upper, middle and lower slopes (T2); and crops were planted along transverse slopes with mulberry trees on the middle slope and on the four sides of the slope (T3). The results showed that soil available nitrogen, available phosphorus, potassium and organic matter contents were significantly improved with mulberry intercropping. The sequence of improvement was T2>T3>T1>CT. The ratios of carbon/nitrogen and carbon/phosphorus changed from 7 to 20 and from 10 to 27 respectively for the four treatments. The carbon/nitrogen and carbon/phosphorus ratios were largest in the T2 treatment for every slope level. Soil organic matter, nitrogen, potassium, carbon/nitrogen ratio, carbon/phosphorus ratio and crop output were all significantly positively correlated. In conclusion, crop-mulberry intercropping in hillside fields improved and maintained soil nutrients and to some extent decreased the ratios of carbon/nitrogen and carbon/phosphorus in uphill soils. These results can be used to guide the choice of water and soil conservation cropping modes in the Three Gorges Reservoir Area.

    Effects of different soil management methods on the soil nutrients, enzyme activity and fruit quality of pear orchards
    WEI Shu-Wei, WANG Shao-Min, ZHANG Yong, RAN Kun, WANG Hong-Wei
    2015, 24(12):  46-55.  DOI: 10.11686/cyxb2015149
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    In order to investigate the effects of different soil management methods on the soil nutrients, enzyme activity and the flavor of ‘Yali’ pear, an experiment has been undertaken with 2 treatments of naturally grown grass and mushroom compost mulching, with clean tillage as the control. Mineral nutrition, the species and quantity of microorganisms, soil enzyme activity and the flavor of Yali pear were measured. Results showed that the organic contents of pear orchard soils treated with mushroom and naturally grown grass mulches increased considerably compared to the control. In orchards with 3 years of naturally grown grass mulching, the organic matter content of 0-20 cm soil was 15.05 g/kg, 1.9 times the control. Alkaline hydrolysis available N, B and Zn were also significantly higher than the control. The contents of available K, Fe, exchangeable Ca and Mg in 0-20 cm and 20-40 cm soil samples were less than the control. For orchards with 3 years of mushroom mulching, the organic matter content of 0-20 cm soil was 19.54 g/kg, 2.4 times the control. Available N, P, K, Fe, Zn, boron, exchangeable Ca and Mg in 0-20 cm and 20-40 cm soil samples were also higher than the control. Naturally grown grass and mushroom compost mulches improved the quantity and activity of soil microorganisms. In pear orchards with 3 years of naturally grown grass treatment, the quantities of bacteria, fungi and actinommyces in 0-20 cm soil was 163.40%, 18.75% and 166.24% higher than the control. In orchards with 3 years mushroom mulching, they were 402.33%, 37.50% and 163.05% higher. Phosphatase activities under naturally grown grass and mushroom mulching were 1.45 and 2.00 times higher than the control, while urease activity was 1.94 and 2.39 times higher. Naturally grown grass and mushroom compost mulching significantly enhanced the quality of fruit flavors. In both cases, the range of aromas and sugar contents of ‘Yali’ pears were significantly higher than the control.

    Influence of crop rotation on soil nutrients, microbial activities and bacterial community structures
    CHEN Dan-Mei, CHEN Xiao-Ming, LIANG Yong-Jiang, HUO Xin-Jian, ZHANG Chang-Hua, DUAN Yu-Qi, YANG Yu-Hong, YUAN Ling
    2015, 24(12):  56-65.  DOI: 10.11686/cyxb2015002
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    A 16-year field experiment has been carried out to investigate the influence of crop rotations on soil nutrients, microbial activities and bacterial community structures. The study includes 454 pyrosequencing analysis of bacterial 16S rDNA. The experiment investigated four crop rotation treatments: 1) flue-cured tobacco in summer, followed by winter fallowing and maize in summer (T-F-M); 2) flue-cured tobacco in summer, canola in winter and maize in summer (T-C-M); 3) flue-cured tobacco in summer, canola in winter and rice in summer (T-C-R); and 4) tobacco in summer, Vicia villosa in winter and rice in summer (T-V-R). After 16-years of crop rotation, soil pH had changed from 6.2 to 7.1. Compared with original soil levels, organic matter and available nutrients (such as nitrogen, phosphorus and potassium) either varied very little or significantly increased, suggesting the retention or growth of fertility under intensive rotations that return straw into soil and fallow during winter. There were higher microbial biomass carbon and nitrogen and enzyme activity (for example, sucrase, phosphatase, dehydrogenase, catalase and urease) in soils under the T-V-R treatment than in any of the other treatments. The readings of bacterial 16S rDNA sequences were 5395 for T-F-M soils, 5864 for T-C-M, 5528 for T-C-R and 6614 for T-V-R, representing 885, 973, 969 and 987 bacterial genera or species respectively, attributed to 31 phyla including Proteobacteria, Chloroflexi, Actinobacteria, Acidobacteria and Bacteroidetes. Proteobacteria were the largest group, accounting for 24.69%-32.49% of soil bacteria. 5 of the 31 bacterial phyla and 6 of the 15 predominant bacteria were found in all the treatments’ soils. It seems reasonable to suggest that crop rotation was a fundamental determinant of the abundance of soil bacterial components and community structures. Taking into account variations in organic matter, available nutrients, enzyme activities, microbial biomass, bacterial community structures and biodiversity, our experiment suggests that T-V-R could be the best rotation model to promote in the flue-tobacco cultivation areas of Yunnan province.
    Microbial characteristics of Hordeum brevisubulatum silage and screening for lactic acid bacteria with high fermentation performance
    TAO Ya, LI Feng, GAO Feng-Qin, SUN Qi-Zhong
    2015, 24(12):  66-73.  DOI: 10.11686/cyxb2015138
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    A study has been undertaken to investigate the microbial community structure, microorganism populations and diversity of lactic acid bacteria in Hordeum brevisubulatum silage. Lactic acid bacteria, coliform bacteria, aerobic bacteria, yeast and mold were isolated and counted by means of selective media. Lactic acid bacteria strains were identified by morphological observation, physiological and biochemical characteristics and partial 16S rDNA gene sequences. In order to assist the development of beneficial microorganisms for animal feed, the lactic acid bacteria strains with high fermentation performance were screened based on growth curves and their ability to produce and resist acid. The results showed that microorganism populations change in different ways through the fermentation process. Lactic acid bacteria populations increased from 0 to 4.00×108 cfu/g FM and yeast populations grew from 8.50×105 to 1.02×108 cfu/g FM, whereas coliform bacteria, aerobic bacteria and mold populations did not change significantly. Four lactic acid bacteria strains were isolated from H. brevisubulatum silage which were identified at Lx36 isolation as Lactobacillus pentosus, at Lx37 isolation as L. brevis, at Lx53 isolation as Pediococcus pentosaceus and at Lx54 isolation as L. parabuchneri. Lx36 was screened out for high fermentation performance because it ceased to grow after 20 hours of cultivation, with an OD value of 4.21. The pH value of MRS broth (the bacterial growth medium) was only 4.08 after 12 hours of Lx36 cultivation, and the strain can survive in MRS broth with a pH of 3.0. These results indicate that silage fermentation is a process of microorganism interaction and that silage quality is directly influenced by the changing character of microorganism populations. Lactic acid bacteria species in H. brevisubulatum silage were relatively abundant. L. pentosus screened with a high ability to reproduce and to produce and resist acid, suggesting its potential value for practical applications.
    Pathogenic mechanism of Rhizoctonia solani potato blight Ⅰ Micro-structure observation of the infection process and measurement of cell wall degradation enzyme activity
    TUO Ning, ZHANG Jun, QIU Hui-Zhen, ZHANG Wen-Ming, ZHANG Chun-Hong, LIU Xing, ZHU Jing
    2015, 24(12):  74-82.  DOI: 10.11686/cyxb2015051
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    Stem canker (black scurf disease) caused by Rhizoctonia solani has become one of the main soil-borne diseases which limit the development of the potato industry in Gansu province. In this study, R. solani spores were inoculated onto the stem base of potato plants grown in pots. The infection process and activity of potato cell wall degradation enzymes was observed and measured. Results showed that the infection sequence was bud, stem, root, stolon and lastly tuber. Hyphae attached to the surface of the host within 12 h, appressoria formed in 36 h, infection cushions formed in 48 h. Deformation and fracture of the cell wall, damage to the cell membrane, and the effects of plasmolysis such as deformation and fracture of the cell wall, damage to the cell and plastid membranes and the dissolution of cytoplasm were then observed under the electron microscope. The activities of cellulase (Cx), polygalacturonase (PG) and polymethyl-galacturonase (PMG) were 1.7, 2.9 and 3.1 μg/(h·g), respectively, and these values were significantly higher than those of the control group. The activities of PG and PMG were as twice those of the control group. The increased enzyme activity may be related to tissue necrosis.
    Effects of disease control and growth promotion of Polygonum viviparum endophytic bacteria Bacillus mojavensis on potato
    CHANG Tao, YANG Cheng-De, XUE Li, YANG Xiao-Li, FENG Zhong-Hong, HAO Rong-Rong, ZHANG Zhen-Fen, CHEN Xiu-Rong
    2015, 24(12):  83-91.  DOI: 10.11686/cyxb2014233
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    This research was to study the effects of disease prevention, growth promotion and defense enzymes induction of Bacillus mojavensis ZA1 on potato, and provide a theoretical basis for microbial fungicide and fertilizer use. The abilities of IAA secretion, nitrogen fixation, phosphate solubilization and inhibition enzyme production of ZA1 have been researched qualitatively by general methods. The effects of controlling disease and growth promotion of ZA1 on potatoes were studied under the condition of indoors and fields. The concentration of IAA secreted by ZA1 in the King medium with and without tryptophan were 12.17 and 9.75 mg/L. ZA1 possessed the capacity of nitrogen fixation and extracellular proteases, chitinase and glucanase production, but without the ability of phosphate solubilization. The control efficiency of ZA1 was 85.9% by spraying 10 times diluting fermentation broth on potato tubes in storage-period against potato gangrene, and was 26.56% by seed dressing fermentation broth with diluting for 20 times on potato tubes under field condition against potato late blight. In field condition, the production ratios of commodity potato were increased by 36.29% and 33.88% per hectare, respectively. Pot experiments with the seed dressing potatoes showed that the content of roots, stems and chlorophyll were higher than the control group. After treatment by ZA1 20 times fermentation broth on potato tubes, the length of the root and wet and dry weight were increased by 8 cm, 0.75 g and 5.07 g, respectively. In the same time, the plant height, stem diameter, stem wet and dry weight and the content of chlorophyll were increased by 2.74 cm, 0.27 cm, 0.52 g, 5.73 g and 0.54 mg/g, respectively. The root-shoot ratios of wet and dry weight were increased by 0.214 and 0.094, respectively. When spraying diluting fermentation broth of ZA1 on potato leaves, the results indicated that the activity of catalase (CAT), polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL), SOD and POD of potatoes were increased. ZA1 possessed the biological function of disease prevention and growth promotion under indoor and field conditions obviously, which showed that ZA1 had the potential to become microbial fungicide and fertilizer.
    Seasonal evaluation of nutrition and carrying capacity of grazing pastures in the Three-River Source Region
    SUN Peng-Fei, CUI Zhan-Hong, LIU Shu-Jie, CHAI Sha-Tuo, HAO Li-Zhuang, WANG Xun
    2015, 24(12):  92-101.  DOI: 10.11686/cyxb2015024
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    To explore seasonal variations in the potential nutrient supply and carrying capacity of grazing pastures in the Three-River Source Region, three adult Datong yaks equipped the permanent rumen fistula were chosen to provide rumen fluid. Analysis of this fluid was combined with measurement of grass yield to comprehensively determine the pastures’ carrying capacity and gas production, using general nutrient analysis and in vitro techniques. The results showed that the maximum yields of edible dry grass in summer, autumn, winter and spring were (123.83±17.88), (256.88±29.90) and (246.83±66.73) g/m2 respectively. Maximum crude protein (CP) contents were (12.69±0.13)%, (10.54±1.22)% and (8.65±0.64)%, the contents gradually decreasing with grass growth. The maximum ether extract (EE) contents in summer, autumn, winter and spring were (2.95±0.10)%, (4.38±0.17)% and (3.74±0.70)% respectively. The change trend in neutral detergent soluble (NDS) content was consistent with CP, while neutral detergent fibre (NDF) and acid detergent fibre (ADF) contents varied negatively with CP. The pH value and NH3-N concentration of solutions fermented in vitro were within normal ranges. The maxima of 48 h gas production and digestible energy (DE), metabolizable energy (ME) and organic matter degradation rate (OMD) (estimated by 24 h gas production) for summer pastures were (57.50±4.27) mL, (9.32±0.59) MJ/kg, (7.98±0.62) MJ/kg and (57.93±3.23)%; for autumn pastures (54.67±5.35) mL, (8.83±0.64) MJ/kg, (7.47±0.68) MJ/kg and (55.26±3.52)%; for winter and spring pastures (58.83±4.51) mL, (9.56±0.60) MJ/kg, (8.24±0.63) MJ/kg and (52.69±5.14)%. Without the provision of concentrates, according to grass and digestible crude protein (DCP) production results, the optimal yak carrying capacity of summer, autumn and winter and spring pastures was 7.05,19.51 and 2.47 SU/ha respectively. When supplementary concentrates were fed, maximum carrying capacity for the three seasons can reach to 14.85, 29.00 and 5.03 SU/ha (based on DCP production for summer pastures and ME production for autumn, winter and spring pastures). In conclusion, the forage yield and quality of grazing pastures varies significantly by season. By supplementary feeding, carrying capacity in summer, autumn, winter and spring can be increased by 1.1, 0.5 and 1.0 times, a result that would help to promote both local animal husbandry and ecological sustainability.
    Effects of different energy levels and corn processing diets on ruminal fermentation parameters in vitro
    ZHANG Ting, ZHANG Bin, ZHANG Pei-Hua, ZHOU Xiao-Qiao, TIAN Yao, ZHU Dan, ZHAO Meng, LIU Shi-Jie, ZHANG Kai-Zhan, CHEN Yu-Guang, BU Deng-Pan, William P.Weiss
    2015, 24(12):  102-111.  DOI: 10.11686/cyxb2015060
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    In vitro batch cultures were grown to investigate the effect of different energy levels and corn processing diets on ruminal fermentation parameters in a 2×2 factorial experiment. Isonitrogenous diets were constructed with a concentrate-to-forage ratio of 65∶35 using corn straw as the main forage, and corn silage and rumen-protected fat were added to adjust the diets’ energy levels. In addition, steam-flaked corn and ground corn were tested as different corn processing products. The results showed that the theoretical maximum gas production, 48 h total gas production and dry matter degradability of the high-energy diets were significantly higher than for the low energy diets (P<0.05). The acetate and propionate of high-energy diets were significantly higher than low-energy diets with ground corn (P<0.05). At the same energy level, the 48 h total gas production of diets with steam-flaked corn was higher than for ground corn (P<0.05). Furthermore, at the low-energy level the total volatile fatty acid and propionate of diets with steam-flaked corn were higher than for ground corn (P<0.05). Correlation analysis between the different feed nutrition levels and fermentation parameters in vitro showed that 48 h total gas production was significantly negatively related to neutral detergent fiber (P<0.05), while it had a significantly positive correlation with neutral detergent-soluble (NDS) and non-fiber carbohydrates and with NDS/crude protein (P<0.05). In conclusion, both steam-flaked corn and high-energy level diets can increase 48 h total gas production in vitro, and steam-flaked corn can improve ruminal fermentation under low-energy level diets.
    Effect of nitrogen fertilization rate and cutting height on yields, nutritive values and root-rot incidence in forage ramie
    BAI Yu-Chao, GUO Ting, YANG Rui-Fang, SHE Wei, CAO Yi, XIAO Cheng-Xiang, CUI Guo-Xian
    2015, 24(12):  112-120.  DOI: 10.11686/cyxb2015294
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    Ramie (Boehmeria nivea) is rich in nutrients, and is a protein source for animal feed. The objective of this study was to investigate the effects of nitrogen fertilization rate and cutting height on forage yields, nutritive values and root-rot (a common disease in ramie) of Xiangzhu-3 and Tri-1 varieties, and thus provide advisory support for use of Xiangzhu-3 and Tri-1 ramie varieties as a forage crop. A field trial was conducted with a two-factor randomized block design (3×3), including three cutting height levels (40 cm, D1; 70 cm, D2; 100 cm, D3) and three levels of nitrogen fertilizer (0 kg/ha, N1; 92 kg/ha, N2; 138 kg/ha, N3). Dry matter yield (DM), nutrient content and root-rot incidence were measured to elucidate the effect of different nitrogen fertilization and cutting height treatments on the feeding value of Xiangzhu-3 and Tri-1 ramie varieties. Nitrogen fertilization and cutting height had significant effects on biomass, dry matter (DM) yield, crude protein (CP), crude fat (EE), crude ash (Ash), crude fiber (CF) and phosphorus (P) contents of Xiangzhu-3 and Tri-1. Cutting height was the dominant factor affecting DM and CF. For CP content, Xiangzhu-3 was most influenced by cutting height, while Tri-1 responded to by nitrogen fertilization. Both EF and CF data displayed significant interaction between cutting height and nitrogen fertilization. Additionally, both ramie varieties had greatest DM, CP, and EE under the N2 treatment within each cutting height treatment, while the CF content was decreased with increased nitrogen fertilization levels. In each nitrogen fertilization treatment, the CP, calcium (Ca) and P contents of Xiangzhu-3 and Tri-1 were decreased and the biomass and CF were increased with increased cutting heights. Increasing cutting frequency increased the root-rot incidence of Xiangzhu-3 and Tri-1. In summary, this study indicated that the optimal clipping height and nitrogen fertilization treatments of Xiangzhu-3 and Tri-1 ramie varieties were 70-100 cm and 92 kg/ha, respectively.
    Milk production, nitrogen excretion and blood biochemical parameter responses to dietary addition of compound yeast cultures in dairy cows
    WANG Ling, LV Yong-Yan, CHENG Zhi-Wei, DU Gao-Tang, LI Jin-Lin, FU Shi-Jun, SUN Guo-Qiang
    2015, 24(12):  121-130.  DOI: 10.11686/cyxb2015124
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    This experiment was conducted to determine the effects of compound yeast cultures on the milk production, nitrogen excretion and blood biochemical parameters of dairy cows. A total of 24 Holstein lactating cows with similar parity, weight, milk yield and lactation stage (135±15 day in milk) were equally assigned to four groups, including one control group and 3 treatment groups, with dietary supplementation of compound yeast cultures at concentrations of 0, 0.8%, 1.0% and 1.2%, respectively. Milk yield, milk composition, nitrogen excretion and blood biochemical parameters of each cow were recorded. The results showed: 1) The cows in the respective treatment groups had 8.48%, 10.05%, 8.97% higher daily milk yield than cows in the control group (P<0.05). 2) Compound yeast cultures significantly increased milk fat and protein concentration (P<0.05), and reduced somatic cell counts (P<0.05), with treatment group 2 again the greatest response. 3) The supply of compound yeast cultures in dairy cow feed concentrate reduced nitrogen excretion by 8.47%, 12.01%, 9.36% compared to the control group (P<0.05). 4) Dietary supplementation with compound yeast cultures significantly increased the levels of glucose (GLU), total protein (TP), globulin (GLOB), and insulin (INS) (P<0.05) in the serum, and reduced blood urea nitrogen (BUN) (P<0.05). Based on the data for milk yield, milk composition, nitrogen excretion and blood biochemical parameters, the optimal concentration of compound yeast cultures administered as a supplement for dairy cow diets is 1.0%.
    Effects of diet pellets with different concentrate-roughage ratios on rumen fermentation parameters and microorganism abundance in weaned bull calves
    YANG Hong-Bo, LIU Hong, ZHAN Jin-Shun, LIN Miao, ZHAO Guo-Qi
    2015, 24(12):  131-138.  DOI: 10.11686/cyxb2015022
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    This study was conducted to investigate the effects of diet pellets with different concentrate-roughage ratios on rumen microbial protein synthesis, fermentation and microorganism abundance in weaned bull calves. A total of 12 healthy, weaned Holstein bull calves (age=103.92±1.5 d; weight=121.25±4.12 kg) were randomly assigned to 4 groups (Ⅰ, Ⅱ, Ⅲ and Ⅳ), with 3 calves in each group. The treatment diets were complete-diet pellets with four concentrate-roughage ratios (75∶25, Ⅰ; 70∶30, Ⅱ; 65∶35, Ⅲ; 60∶40, Ⅳ). The experiment ran for 70 days in total, including 14 d for adaptation and 56 d for the trial itself. Rumen microbial protein synthesis, fermentation and microbial abundance were measured. The results showed that total volatile fatty acids (VFA) and acetate concentration in group Ⅳ were the lowest, and were significantly lower than those in groupⅠ(P<0.05). The butyrate level of group Ⅳ was significantly higher than groupsⅠand Ⅲ (P<0.05). Group Ⅲ had the highest A∶P, which was significantly greater than group Ⅳ(P<0.05). The relative abundance of Ruminococcus albus, Butyrivibrio fibrisolvens and general fungi in group Ⅳ was significantly higher than the other three groups (P<0.05). Ruminococcus flavefaciens abundance was the highest in group Ⅲ when compared with the other groups (P<0.05). The results indicate that rumen total VFA and acetate concentration can be increased in high-concentrate groups, while it will inhibit growth of the fibre-adherent rumen bacteria and anaerobic fungi.
    Proliferation stimulus and antioxidant effect of alfalfa flavonoids on dairy cow mammary epithelial cells cultured in vitro
    SU Xiao-Shuang, ZHAN Jin-Shun, ZHAN Kang, LIU Ming-Mei, ZHAO Guo-Qi
    2015, 24(12):  139-145.  DOI: 10.11686/cyxb2015030
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    This experiment studied the proliferation stimulus and antioxidant effects of different concentrations of alfalfa flavonoids on cow mammary epithelial cells cultured in vitro. Media with five different concentrations of alfalfa flavonoids were prepared to culture dairy cow mammary epithelial cells in vitro. Concentrations tested were: 0 μg/mL (control group), 25 μg/mL (trial group Ⅰ), 50 μg/mL (trial group Ⅱ), 75 μg/mL (trial group Ⅲ), 100 μg/mL (trial group Ⅳ). The results showed: 1) There was no significant difference in cell proliferation between the five concentrations during the first three days of cell culture, but the cell proliferation of trial groups Ⅱ-Ⅳ was significantly higher than for the control group and trial group Ⅰ (P<0.01) at day 5.2) The NO levels of trial groups Ⅱ-Ⅳ were significantly higher than the control group and trial group Ⅰ (P<0.01). Also, LDH activity of groups Ⅰ, Ⅱ, and Ⅳ was significantly lower than the control group and than test group Ⅲ (P<0.01), and the difference between the control group and test group Ⅲ was not significant. 3) Intracellular CAT content of trial group Ⅳ was significantly higher than the other groups (P<0.01). The MAD content of group Ⅲ was significantly lower than the other groups (P<0.01). GSH-PX activity of groups Ⅱ and Ⅲ was significantly higher than the other groups (P<0.01). In addition, there was no significant difference between the five groups in the cell SOD content. 4) The relative expression levels of p53, Caspase-3, SOCS3 and STAT1 genes in groups Ⅱ and Ⅲ were significantly lower than control group (P<0.01). In summary, alfalfa flavonoids can promote the proliferation of bovine mammary epithelial cells cultured in vitro, improve cell antioxidant capacity and inhibit cellular apoptosis. The best effect occurred at a dosage of 75 μg/mL of alfalfa flavonoids.
    Development of phytomers produced by root crown shoots in Medicago sativa
    FANG Qiang-En, ZHANG Bo, SHI Shang-Li
    2015, 24(12):  146-154.  DOI: 10.11686/cyxb2015253
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    The phytomer, consisting of a node, leaflike organ, internode, and lateral meristem in the axil of a leaf, is a basic structural unit of shoot growth and development. With scanning electron microscopy (SEM), the developmental characteristics of phytomers in alfalfa variety ‘WL168’ were observed throughout the developmental phases of the apical meristem in crown buds from sprouting to flowering. During the development of phytomers, the primordial compound leaf emerged in an acropetal pattern. Both the primordial axillary bud and compound leaf originated from the apical meristem, but they differed in the rhythm of development; phytomers showed two patterns of development. Pattern-I was featured by the faster development of primordial compound leaves than primordial axillary buds which appeared after the formation of lateral primordial leaves. For patternⅡ, primordial axillary buds grew faster than primordial compound leaves, initiating after the formation of primordial stipules. Before the crown bud emerged from the soil, the phytomer developed through pattern-Ⅰ, and immediately after emergence, through pattern-Ⅱ. When the number of aboveground phytomer reached 16-17, the axillary meristem became reproductive. It was concluded that the shift of phytomer development from pattern Ⅰ to pattern Ⅱ can be an indicator of the transition from vegetative to reproductive growth in alfalfa.
    Reproductive organ variation in Fritillaria unibracteata along an altitudinal gradient
    CHEN Wen-Nian, XIAO Xiao-Jun, CHEN Fa-Jun, WANG Hui, ZHANG Zhi-Yong, QI Ze-Min, HUANG Zuo-Xi
    2015, 24(12):  155-163.  DOI: 10.11686/cyxb2015096
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    Three sites (high, medium and low altitude) were established in an alpine mountainous region in the eastern Tibetan Plateau. The reproductive organs of Fritillaria unibracteata at each site were measured. A transplant experiment using F. unibracteata was also carried out; plants from low altitude were transferred to the high site and plants from the high site transferred to the low altitude site. Ecological factors such as snow-melt duration, air temperature, soil water content and soil pH were measured. Reproductive organs including flower, fruit and seed of F. unibracteata were also measured. Snow-melt duration, air temperature and soil water content differed among the different altitudinal sites significantly. Snowpack at the low site melted earlier than those at the other two sites; average air temperature (in March, April and May) at the low site was higher than the other two sites; however soil water content at the low site was the lowest. The highest values for petal length and width, filaments, anthers, styles and style ramifications were measured at the high site and the least at the low site. Flower biomass at the high site was 39.4% heavier than that at the low site, and the ratio of flower biomass to aboveground biomass at the high site was 12.3% higher than that at the low site. Dry weight per fruit at the low site was 48.3% heavier than that at the high site, reaching 1282 mg. Seed number per fruit at the high site was the lowest, 38.1% less than that at the low site. The 100-seed weight at the high site was 142.3 mg, 94.6 mg at the low site. The transplant experiment verified the results described above. We concluded that the reproductive organs of F. unibracteata were significantly affected by altitude factors, particularly snow-melt duration, air temperature (especially sub-zero temperatures), ultraviolet light and insect pollinators.
    Phenotypic variation of wild Arundinella hirta populations from Liaoning province
    LIU Ying, BAI Long, LEI Jia-Jun
    2015, 24(12):  164-179.  DOI: 10.11686/cyxb2015222
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    The phenotypic variation of eight Arudinella hirta populations collected from Liaoning province was determined. Ten phenotypic traits of eight wild populations were analysed using the coefficient of variation, principal component analysis and cluster analysis. There were significant differences in morphological characteristics within and between populations of A. hirta at P<0.01. The coefficients of variation were ranked as follows: total spikelet number/panicle (42.01%)>flag leaf depth (27.94%)>flag leaf length (26.83%)>flag leaf width (20.98%)>leaf number (19.64%)>stem diameter (19.70%)>panicle length (19.30%)>tiller dry weight (18.62%)>plant height (10.19%)>panicle dry weight (8.16%). Principal component analysis based on ten morphological characteristics showed that three principal components reflected most of the phenotypic characteristics; differences between populations were derived from panicle length, panicle dry weight, tiller dry weight and spikelet number. UPGMA cluster analysis identified three groups including tall plants with long and wide flag leaves, tall plants with long and narrow flag leaves and short plants with short and narrow leaves. Correlation analysis showed that some traits were strongly correlation with phenotypic traits and ecological factors.

    Analysis of codon usage in the chloroplast genome of Medicago truncatula
    YANG Guo-Feng, SU Kun-Long, ZHAO Yi-Ran, SONG Zhi-Bin, SUN Juan
    2015, 24(12):  171-179.  DOI: 10.11686/cyxb2015016
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    The complete nucleotide sequence of the chloroplast genome of Medicago truncatula was investigated. Fifty CDS (coding DNA sequences) selected from the chloroplast genome sequence of M. truncatula, were analyzed using CodonW software. The results show that the third codon position was rich in A and U. ENC ranged from 37.1 to 51.9 meaning that the codon bias was weak. There were 23 codons with relative synonymous codon usage greater than 1 and 20 codons ending with A and T. ENC-plot analysis showed that GC3 was not correlated with GC12; ENC ratio’s of most genes ranged from -0.05 to 0.05. In the correspondence analysis of the first group of four axes, the first axis showed 10.3% variation. The correlation coefficients for axis 1 between ENC and GC3 were 0.091 and -0.092 respectively (not significant). Synonymous codon usage bias was found, mainly due to the effect of mutation pressure, but there were other factors. In addition, analysis of the high expression codons enabled 23 to be affirmed as the “optimal codons” as UAA, UUG, CCU. The results provide evidence for molecular modification of exogenous genes to increase the expression efficiency in M. truncatula chloroplasts.
    Relationship between dormancy breaking and protective enzymes in refrigerated Lilium pumilum bulbs
    LIU Fang, LI Dan-Dan, LIAN Hua, WU Xia, CHEN Ye-Wen, LI Yu-Qi, XIE Shou-Feng, CHEN Xue-Wu
    2015, 24(12):  180-187.  DOI: 10.11686/cyxb2015031
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    Changes in bulb protective enzyme activity and carbohydrate content of Lilium pumilum bulbs held at 5℃to break dormancy were investigated. Results showed that superoxide dismutase (SOD) activity of scales, apical buds and basal plates decreased within 0-24 days (d). Except for apical buds, peroxidase (POD) activity within 0-24 d declined in all bulb parts and catalase (CAT) activity of the outer scales, apical buds and basal plates decreased within 0-12 d. During middle and later periods, SOD, CAT and ascorbate peroxidase (ASP) increased, POD activity decreased, POD and polyphenol oxidase (PPO) activity of apical buds and basal plates also increased in the middle of the cold storage period. The roles of various metabolism related enzymes in different organs were not the same. With low temperature treatment for 36-60 d, ASP and CAT activity initially rapidly increased but later stabilized. The minimum value of SOD activity was found after 36-60 d cold storage. Starch content was negatively correlated with CAT, PPO, ASP, SOD and POD in all bulb parts; phenylalanine ammonia lyase (PAL) was positively correlated with starch in the scales. Bulb dormancy breaking started at 36 d, and was mostly complete at 60 d.
    Isolation, identification and biological activities of alkaloids from Anisodus tanguticus
    WANG Yu-Ling, HU Guan-Fang, LIU Min-Yan, YU Hai-Tao, NIU Shu-Jun, LI Yu-Qi
    2015, 24(12):  188-195.  DOI: 10.11686/cyxb2015057
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    Two alkaloids from the aerial parts of Anisodus tanguticus were isolated by elution through series of gel columns. The alkaloid chemical structures were determined by MS and NMR and the alkaloids were identified as hyoscyamine and scopolamine . Contact toxicities of the two alkaloids on nine species of aphids including Brevicoryne brassicae and Myzus persicae, and four species of mites including Tetranychus cinnabarinus and T. urticae were measured by topical application and slide dip methods. The results showed that hyoscyamine had higher contact toxicity than scopolamine on Rhopalosiphum padi, Hyalopterus arundimis, Aphis craccivora and A. citricola with LC50 values of 257.863 , 275.459, 344.645 and 344.717 mg/L, respectively. However, scopolamine was more effective in controlling A. craccivora and R. padi with LC50 values of 311.585 and 392.309 mg/L, respectively. The control pesticide rotenone was most toxic to B. brassicae and Aphis gossypii with LC50 values of 399.542 and 436.124 mg/L, respectively. Contact toxicities of hyoscyamine on the nine species of aphids including B. brassicae were higher than or similar to scopolamine. Compared to rotenone, hyoscyamine and scopolamine were more effective against A. craccivora and R. padi. However, rotenone was more effective against B. brassicae, M. persicae and A. gossypii. For the four species of mites tested (T. truncates, T. viennensis, T. urticae and T. cinnabarinus), hyoscyamine and scopolamine had similar effects, which included higher contact toxicities on T. urticae and T. cinnabarinus (LC50: 201.027-224.172 mg/L) than on T. truncates and T. viennensis (LC50: 257.014-332.698 mg/L). Rotenone was more effective than the two alkaloids against T. truncates and T. viennensis with LC50 values of 196.847 and 224.592 mg/L, respectively.
    Effects of NaCl on seed germination and seedling growth of Halocnemum strobilaceum
    FANG Juan-Juan, WU Yu-Ling, LIU Yun, GONG Hui-Fang, MA Rong-Gang, Hu Ling-Zhi, CHEN Hui
    2015, 24(12):  196-203.  DOI: 10.11686/cyxb2015058
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    In order to identify the optimal NaCl concentration in MS (Murashige and Skoog) medium for seed germination and growth of Halocnemum strobilaceum, and to reveal the primary mechanism of salt osmotic stress (using MS salt medium) promotion of root growth in this species. We used MS medium as a basic medium and added different concentrations of NaCl (0-700 mmol/L) and measured the response of seed germination and seedling growth by culturing seeds stored for 7 years. In addition seedling root tips were cultivated on media with optimum salt concentration to allow morphological and cytological observations under a light microscope. After 7 years of storage seed retained vigor; the seed vigor index and seed germination rate of the control treatment was 3.36 and 74.46% respectively, but germination time appeared to be delayed. Seed germination was not synchronized, beginning after 6-8 days and continued for 25-30 days. The optimum concentration of NaCl in MS medium for seed germination ranged from 100 to 200 mmol/L and compared with the control group, seeds germinated 2 days earlier. However, at NaCl concentrations over 400 mmol/L seed germination was strongly inhibited. The most appropriate salt concentration range in MS medium for seedling growth was 100-300 mmol/L and the optimum concentration was 200 mmol/L. Observation of plant phenotype revealed that seedlings cultured on media with no or medium salt (>300 mmol/L) produced red hypocotyls and subsequently red pigmentation in most plant tissue (3-4 months growth). Root tips from seedlings cultured in medium salt conditions had accelerated root tip cell division and elongation and subsequently promoted root cap cell death and abscission.This investigation provided a good foundation for further study on the salt resistance mechanism of H. strobilaceum.
    Impacts of salt stress on the growth and physiological characteristics of Paspalum vaginatum
    JIA Xin-Ping, DENG Yan-Ming, SUN Xiao-Bo, LIANG Li-Jian
    2015, 24(12):  204-212.  DOI: 10.11686/cyxb2015028
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    The effects of different concentrations of NaCl solution (0, 100, 200, 300, 400 and 500 mmol/L) on growth, biomass, relative water content, membrane permeability, photosynthetic pigments, organic osmolyte content and protective enzyme activity of Paspalum vaginatum were assessed to determine the response of growth and physiological parameters to salt stress. The results indicated that plant height, leaf length, leaf width, root dry weight, stem dry weight, leaf dry weight and erect stem diameter of P. vaginatum under NaCl treatment were significantly reduced compared with the control (no salt). Compared with the control, the relative water content of P. vaginatum leaves was decreased significantly, while the membrane permeability, photosynthetic pigment, MDA and soluble sugar content increased. The activities of SOD and POD and leaf proline content all increased at lower salt levels and then declined, peaking at 300 mmol/L NaCl. This study showed that P. vaginatum has self protecting mechanisms providing tolerance to salinity stress; the threshold of salt tolerance was 300 mmol/L NaCl. The improvement of salt-tolerance in P. vaginatum resulted from the modulation of physiological characteristics.
    Comparative analysis of endogenous hormones in maize seeds with different dormancy characteristics
    WU Yuan-Qi, LENG Yi-Feng, XIA Chao, ZHOU Shu-Feng, LAN Hai
    2015, 24(12):  213-219.  DOI: 10.11686/cyxb2015136
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    A study has been undertaken to clarify maize seed dormancy mechanisms. Four endogenous hormones-zeatin (ZT), gibberellins (GA3), indole acetic acid (IAA) and abscisic acid (ABA) in 7 maize inbred lines with diverse dormancy characteristics were measured and analysed by HPLC. We analysed the effect of the four endogenous hormones on seed dormancy, including their interaction. Results indicated that ZT content was negatively correlated with seed dormancy, with an antagonism to germination inhibitors. GA3 is necessary for germination and can help break dormancy. IAA has no correlation with seed dormancy. ABA promoted dormancy and inhibited germination, with an antagonism to GA3. Furthermore, seed dormancy was closely related to the proportion of the four hormones. Maize inbred lines with strong dormancy had the following traits: low content of ZT and GA3, high content of ABA, low values of ZT/ABA and GA3/ABA, and balanced hormone content.
    Research advances in higher plant adaptation to salt stress
    ZHANG Jin-Lin, LI Hui-Ru, GUO Shu-Yuan, WANG Suo-Min, SHI Hua-Zhong, HAN Qing-Qing, BAO Ai-Ke, MA Qing
    2015, 24(12):  220-236.  DOI: 10.11686/cyxb2015233
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    Soil salinity is a serious worldwide problem causing reduction in crop growth and agricultural output potential. Consequently, finding new ways to minimize the adverse effects of soil salinization on agriculture is globally important. Understanding the adaptation mechanisms of higher plants to salt stress is critical for enhancing salt tolerance and yields of crop plants as well as protecting ecological environments. In this paper, we reviewed the key progresses in salt stress adaptation of higher plants, including the effects of salt stress in plants; physiological mechanism of plant salt tolerance (osmotic adjustment, nutrient balance and the antioxidant system); the diversity of genes relevant to salt tolerance (ion transporting protein genes, osmotic regulation-related genes, signal transduction-related genes and cellular antioxidant-related genes and so on); and the approaches for crop improvement in salt tolerance. Prospects for developing crop plants tolerant to salinity are also discussed.
    Evaluation of the adaptability of bioenergy grasses in acidic red soil
    HOU Wei, XIAO Liang, YI Zi-Li, QIN Jing-Ping, YANG Sai, ZHENG Cheng, CHEN Zhi-Yong
    2015, 24(12):  237-244.  DOI: 10.11686/cyxb2015005
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    Seven grass species grown as bioenergy crops were compared using 20 traits including phenological period, winter survival rate, agronomic traits and chemical composition. The results showed that, with the exception of Pennisetum purpureum, all grasses could complete their growth period and grow well in acidic red soil. There were significant differences among the grasses for seven agronomic characters and five chemical composition traits. Miscanthus floridulus, M. lutarioriparia×sinensis and Panicum virgatum had higher dry matter yield than other species; 5385.24, 4846.09 and 4566.83 g respectively. Cellulose contents of Saccharum arundinaceu, Miscanthus×giganteus and M. lutarioriparia×sinensis were 46.13%, 45.38% and 45.16% respectively, significantly higher than other species. Principal component analysis of twelve quantitative traits revealed that the cumulative contribution rate of four principal component factors accounted for was 92.626% of variation. This indicated that M. lutarioriparia×sinensis was the most adaptable species followed by M. floridulus, P. purpureum, S. arundinaceum, P. virgatum, Miscanthus×giganteus and M. sacchariflora. The results of this study will provide a theoretical basis for the development and utilization of acidic red soil in southern China.
    Photosynthetic characteristics of Chlorophytum capense var. medio-pictum under short duration high light intensity
    DONG Li-Hua, HAN Qiao-Hong, YANG Yong, YUAN Ming
    2015, 24(12):  245-252.  DOI: 10.11686/cyxb2015059
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    It is well known that high light often causes decreased chlorophyll in plants, but the photosynthetic and physiological characteristics of reduced chlorophyll variants under high light is not well understood. Physiological responses to high light intensity in the green stripe and yellow stripe leaf variants of Chlorophytum capense var. medio-pictum were investigated in this study. The results showed that under normal light levels, PSⅡ activity of yellow stripe plants was lower than those of green stripe. However, the level of reactive oxygen species remained elevated. After 6 hours of high light stress, the level of reactive oxygen species and activity of antioxidant enzymes varied less in yellow stripe plants. Under a few minutes of high light stress, non-photochemical quenching increased in all tissues, but the non-photochemical quenching of the yellow stripe plants increased less than the others. These results demonstrated that excess light energy in yellow-stripe leaf tissue was not dissipated in the form of heat; furthermore, the level of non-photochemical quenching in yellow stripe plants was restored in a few minutes.