Scientia Agricultura Sinica

Spatial Distribution Regulation of Protein Content of Naked Barley Varieties and Its Relationships with Environmental Factors in Qinghai-Tibet Plateau

WANG JianLin, ZHONG ZhiMing, FENG XiBo, FU Gang, HOU WeiHai, WANG GaiHua, Da-cizhuoga

Scientia Agricultura Sinica. 2017, 50(6): 969-977. doi:10.3864/j.issn.0578-1752.2017.06.001

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【Objective】 The objective of this study is to reveal the effect of different environmental factors on grain protein content (GPC) of naked barley, to understand the relationship between naked barley GPC distribution characteristics and environmental factors, and to get clear on GPC acclimation of naked barley varieties from different areas to the environment of the Qinghai-Tibet plateau. 【Method】 Based on the method of agronomy combined with geography, naked barley GPC distribution characteristics in Qinghai-Tibet plateau was studied. 【Result】 Along the horizontal direction in Qinghai-Tibet plateau, GPC spatial distribution characteristics of naked barley varieties appeared in a macrocosm trend of high-in-south and low-in-north with a staggered pattern, and there developed two regions of higher GPC. One of the regions of higher GPC exists between the area with longitudes from 100.0°E to 102.5°E and with latitudes from 35.0°N to 37.5°N. And the average GPC of barley varieties in this region is (13.1163±0.5939)% in north-eastern Qinghai-Tibet plateau with distribution centers including Gonghe County, Guide County, Menyuan County and Tongde County in Qinghai Province, and Hezuo County in Gansu Province. Another region of higher GPC existed between the area with longitudes from 86.0°E to 92.0°E and with latitudes from 28.0°N to 29.0°N. And the average GPC of barley varieties in this region is (12.8715±0.6609)% in south-central Qinghai-Tibet plateau with distribution centers including Gongga County, Lazi County, Nimu County, Zhanang County, Nielamu County, Duilongdeiqing County, Sangri County and Kangma County in Tibet. Along the vertical direction with altitudes, GPC distribution pattern appeared in shape of inverted letter N. The average GPC value is (10.8650±1.8600)% at the altitudes below 3 000 m. GPC gradually decreased with altitude increasing, and then reached to the lower value at the altitudes from 3 000 m to 3 300 m. And GPC gradually increased with altitude increasing at the altitudes above from 3 000 m to 3 300 m, and then reached to the highest value at the altitudes from 3 600 m to 3 900 m with average GPC value of (10.8937±2.0719)%. Hereafter, GPC of barley varieties decreased gradually with altitude increasing. The magnitude of effect of environmental factors on naked barley GPC follows an order of soil available N content > sunshine hours during the growth period from heading to maturation > average diurnal temperature range during the growth period from seedling from tiller > average diurnal temperature range during the growth period from tiller to elongation > relative humidity during the growth period from elongation to heading. 【Conclusion】 Soil factors are the main environmental factors with the greatest impact on GPC, and followed by climatic factors, the effect of geographical factors on GPC is not apparent. Soil available N content is the main soil factor influencing barley GPC, and sunshine hours during the growth period from heading to maturation, average diurnal temperature range during the growth period from seedling from tiller, average diurnal temperature range during the growth period from tiller to elongation and relative humidity during the growth period from elongation to heading are the main climatic factors. Barley GPC increased with the increase of average diurnal temperature range during the growth period from tiller to elongation and relative humidity during the growth period from elongation to heading, though decreased significantly with the increase of sunshine hours during the growth period from heading to maturation and average diurnal temperature range during the growth period from seedling to tiller and soil available N content.

Heterosis Analysis of Internode Length Under Ear in Maize

LI HuiMin, LI WeiHua, GUO HaiPing, LIU Kun, ZHANG XiangGe, ZHANG XiaoXiang, XIE HuiLing, TANG JiHua, DING Dong

Scientia Agricultura Sinica. 2017, 50(6): 978-989. doi:10.3864/j.issn.0578-1752.2017.06.002

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【Objective】The under-ear internode length determines maize seedling height and ear height, which are two agronomic traits associated with yield and lodging resistance. Heterosis, a wild-spread genetic phenomenon, is widely applied in crop yield and quality improvement. In a previous study, the lengths of the 7th, 8th and 9th internodes were found to have a decisive effect on ear height and showed a high degree of heterosis. The aim of this study is to resolve the molecular mechanism of heterosis for comprehensive understanding and utilization of heterosis.【Method】 In this study, the chromosome single segment substitution lines (Single segment substitution lines, SSSLs) derived from continually cross of lx9801 and Chang7-2 were used as the basic materials. Test-crossing populations were constructed by crossing the SSSLs with inbred lines Zheng58 and Xun9058, respectively. The HL(Heterosis loci) of the 7th, 8th, and 9th internode length were detected through two-year and two-point tests.【Result】By using the SSSL×Zheng58 test-cross group and SSSL× Xun9058 test-cross group, the heterosis loci (HL) of the 7th, 8th, and 9th internode length were detected through two-year and two-point tests. In 2012, the mid parent heterosis values of the 7th, 8th, and 9th internode length in Xunxian were 57.25% and 78.16%, 68.30% and 75.04%, 59.48% and 62.85%, respectively, and that in Changge were 48.27% and 63.02%, 43.36% and 54.80%, 37.26% and 42.62%, respectively. In 2013, the mid parent heterosis values of the 7th, 8th, and 9th internode length in Xunxian were 23.01% and 37%, 22.69% and 35.65%, 22.20% and 34.74%, respectively, and that in Changge were 21.86% and 33.19%, 20.99% and 35.57%, 27.55% and 42.19%, respectively. A total of 18 and 18 length heterosis loci were obtained from the 7th internode, 20 and 23 were obtained from the 8th internode, 17 and 19 were obtained from the 9th internode. Compared the same HL of the two test-crossing groups, there are 3, 3 and 1 same HL of 7th, 8th, 9th internode length, respectively. These same HL occupied 12.7% and 11.6% of the total HL numbers, respectively. 【Conclusion】There were only 7 (6%) same HL sites had same location between SSSL×Zheng58 group and SSSL×Xun9058 group of the 7th, 8th, and 9th internode length, which indicated that the heterotic loci among different groups may be different. It was speculated that under different genetic backgrounds there are different genes controlling the same heterosis traits. The performance of heterotic loci at single gene level should be crossing group (genetic background) specific.

Advances and Perspectives in Research of Potato Genetics and Breeding

XU JianFei, JIN LiPing

Scientia Agricultura Sinica. 2017, 50(6): 990-1015. doi:10.3864/j.issn.0578-1752.2017.06.003

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Potato, the third most important food crop, plays a key role in global and China’s food security. Improvement of varieties is a base for sustainable development of potato industry. Potatoes frequently suffer from diverse biotic and abiotic stress, so it is urgent to breed new varieties with better disease resistance, stress tolerance, tuber yield and quality as well as specific usage to meet the needs of potato processing and people nutrition. Potato breeding is a system combining germplasm evaluation and utilization, major traits genetics analysis, breeding technology application and variety extension and crop management together. Within a global conservation strategy there are about 65,000 accessions. Using a homozygous doubled-monoploid potato clone, 86% of the 844-megabase genome sequence are revealed and assembled, and 39,031 protein-coding genes are predicted. At present, re-sequencing of potato accessions is in process. Common cultivated potato is an asexual propagation tetraploid with tetrasomic inheritance and high heterozygosity. Nevertheless, inheritance of many major traits involving plant development and morphology, tuber quality, disease resistance and stress tolerance are revealed. A lot of genes determining potato major traits are mapped and cloned. Potato breeding technology involves conventional breeding, ploidy manipulation, marker-assisted selection, genetic engineering and promising genomic selection for complex traits. Since 1949, China potato breeding has achieved great progress that is reflected on growth of number of registered varieties. Dutch potato breeding ranks among the best in world and participatory potato breeding model is a successful practice for commercial breeding. In the future, it is a trend to breed superior and specific purpose varieties based on improvement of integrated breeding technology, innovation of breeding model and germplasm utilization.

Effects of Long-Term Tillage and Fertilization on Yield and Water Use Efficiency of Winter Wheat in Loess Dry Land Plateau

ZHANG JianJun, FAN TingLu, DANG Yi, ZHAO Gang, WANG Lei, LI ShangZhong, WANG ShuYing, WANG Yong

Scientia Agricultura Sinica. 2017, 50(6): 1016-1030. doi:10.3864/j.issn.0578-1752.2017.06.004

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【Objective】The aim of this experiment is to investigate the effects of tillage and fertilization on yield and water use efficiency of winter wheat under wheat-spring maize rotation at fields in the Loess Plateau of Eastern Gansu.【Method】The work is based on a 12 year field experiment of tillage and fertilization in Zhenyuan, Gansu province, where is a semi-humid arid area. A split plot design was developed in this experiment, with different tillage systems (traditional tillage and no-tillage) as main treatments and different fertilizer application (CK, N, P, M, NP, NMP) as sub-treatments. The yield and water use efficiency of winter wheat under different treatments were measured, and the relationship between yield and water consumption of winter wheat were further studied.【Result】The results showed that the treatments with organic manure had the higher yield of winter wheat in the same tillage system, implying that organic manure was superior to chemical fertilizer alone in the experimental area. The yield under nitrogen fertilization was lower than phosphate fertilizer. The yield of winter wheat under traditional tillage was higher than that of no-tillage. In terms of water use efficiency, water use efficiency under NMP treatment was the highest among different fertilization methods in different years. The average water use efficiency was in the order of NMP＞NP＞M＞P＞CK＞N; the average water use efficiency in treatment of the combination of NMP fertilizer and traditional tillage increased by 84.0% and that of no-tillage increased by 84.1%. The average water use efficiency under traditional tillage was higher than no-tillage, and that increased by 13.6% in the combination of traditional tillage and NMP fertilizer compared with no-tillage. The water consumption of winter wheat was strongly related to rainfall, and the total water consumption by crops under NMP was the highest. The average annual water consumption of NMP was 3.0%, 4.4%, 31.4% higher than that of CK under traditional tillage in three years; 10.2%, 1.5% and 25.7% were increased, respectively, under no-tillage in three years. On the other hand, NMP treatment significantly reduced the water consumption and the ratio of total water consumption during sowing-greening period and increased the water consumption during the greening-mature period. The total water consumption under the traditional tillage system was higher than under the no-tillage system in the drought year, but the no-tillage system had higher total water consumption in the wet year than in the traditional farming year. No consistent trend was found in rainfall year. In addition, regardless of the kind of rainfall year, different tillage and fertilization treatment, water content showed the change point at about 60 cm soil layer, but this point in NMP was lower than CK and other fertilization treatments, but the order of influence of soil water content was wet year＞normal year＞dry year. In the order of change for the abundance and the marginal water use efficiency, NMP treatment had the highest water use efficiency while as for tillage, traditional tillage had significant results.【Conclusion】The yield, water use efficiency of winter wheat under NMP treatment was the highest in different cropping patterns and rainfall years, where the water content showed the lowest at 60 cm soil depth. Besides, the total water consumption under NMP treatment was relatively higher; which might be helpful to reduce the water consumption and the proportion of water consumption at the stage of greening, and increase the consumption of water consumption and the storage of 0-200 cm soil layer. The abundant water use efficiency and marginal water use efficiency of winter wheat were higher under traditional tillage than that of no-tillage. Therefore, considering the winter wheat yield, water use efficiency and water consumption characteristics, regardless of what kind of rainfall years, the use of traditional tillage with organic and inorganic fertilizer under the conditions of this experiment is the best farming.

Effects of the Year-Round Management Model of N, P and K Combined Application on Grain Yield and Nutrient Efficiency of Wheat-Maize Rotation System in Lime Concretion Black Soil

WANG YongHua, HUANG Yuan, XIN MingHua, YUAN ShaSha, KANG GuoZhang, FENG Wei, XIE YingXin, ZHU YunJi, GUO TianCai

Scientia Agricultura Sinica. 2017, 50(6): 1031-1046. doi:10.3864/j.issn.0578-1752.2017.06.005

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【Objective】This research was conducted to explore the effect of different year-round management models of combined application of nitrogen (N), phosphorus (P) and potassium (K) nutrients on grain yields and nutrient efficiency of wheat-maize rotation system in lime concretion black soil and identify an optimal model for the use efficiency of the combined N, P, and K fertilizers in integrated wheat and maize planting system in the lime concretion black soil in the southeast area of Henan province.【Method】A field experiment of combined application of N, P, and K fertilizers was persistently performed in a typical lime concretion black soil area located in Shangshui country, Zhoukou city of Henan province during two continuous wheat-maize growing seasons (2012.09-2013.08 and 2013.09-2014.08). In this experiment, a wheat-maize rotation system was designed, in which the amounts of total P and K fertilizers were not changed while two different amounts of N fertilizers and different application methods of P and K fertilizers were applied. And the effects of this rotation on absorbance and utilization of N, P, and K fertilizers by wheat and maize crops were studies. In this study, two different amounts of N fertilizers were applied at two levels of 360.00 kg·hm^-2 and 540.00 kg·hm^-2 in every year. The amounts of total P and K fertilizers were not changed, whereas these two fertilizers were applied with four different ways: all P and K fertilizers were applied in wheat and maize growing seasons, respectively; all P fertilizers were applied in wheat growing seasons, and K fertilizers were applied in both wheat and maize seasons and the ratio of K fertilizer applied in these two seasons was 42:58; all K fertilizers were applied at maize growing season and P fertilizers were applied in both wheat and maize seasons with 64:36 ratio for these two crops; P and K fertilizers were simultaneously applied into soil in both wheat and maize seasons and the ratios of P and K fertilizers applied in these two seasons were 64:36 and 42:58, respectively. The aim of this study is to find an optimal combined mode of application of N, P and K fertilizers in wheat/maize rotation system in the lime concretion black soil.【Result】Results of the experiment indicated that under large amount of the applied N fertilizer condition, simultaneous application of P and K fertilizers in both wheat and maize crops could promote the coordinated development of three key components of wheat and maize yields, and significantly improved the spike number of wheat and the lengths as well as row grains of maize spikes. Of all treatments in these two continuous crop growing seasons, P8 treatment (a larger amount of P fertilizer applied to wheat and a larger amount of K fertilizer applied to maize) harvested the highest grain yields in both a single or two seasons, and year-round yields were respectively 21 274.2 kg·hm^-2 and 20 219.1 kg·hm^-2 in two growing seasons (2012.09-2013.08 and 2013.09-2014.08). The nutrient contents in aboveground tissues of wheat and maize crops showed N>K>P in lime concretion black soil. Compared with low amounts of the applied N fertilizer condition, large amounts of the applied N fertilizers were helpful to improvement of the contents of total N, P and K nutrients in the aboveground tissues of these two crops, whereas reduced nitrogen partial factor productivity (NPFP), nitrogen uptake efficiency (NUPE) and nitrogen use efficiency (NUE). Simultaneous application of P and K fertilizers to both wheat and maize crops in one growing year improved the absorbance of N fertilizers, prevented the loss of major nutrients, improved the absorption and utilization of P and K fertilizers of both wheat and maize crops, and significantly increased partial productivity (PFP) and uptake efficiency (UPE) of P and K fertilizers. However, application of total P or K fertilizers in single wheat or maize resulted in the reduced use efficiency of major nutrients and the decreased yields of the subsequently planted crops for unbalanced nutrient supply and the reduced absorbance of N, P and K fertilizers, because P fertilizer is easily immobilized into soil and K fertilizer is easily washed away from soil, respectively. The yield of subsequently planted crops would decrease in this way, because the crops could not absorb and utilize N, P and K in the soil environment of unbalanced nutrient supply. Under small amount of the applied N fertilizer condition, absorbance and utilization of N, P and K nutrients in P4 treatment (a larger amount of P fertilizer applied to wheat and a larger amount of K fertilizer applied to maize) were the highest among all treatments. Under large amount of the applied N fertilizer condition, however, NPFP of N fertilizer in single season or year-round in P8 treatment was the highest among all treatments.【Conclusion】 Based on the efficiency of nutrient utilization and yields of wheat and maize crops, a larger amount of P and K fertilizers separately applied in wheat and maize crops could help synchronously to improve the utilization efficiency of three main nutrients and yields of wheat and maize crops. This can be an optimal model of main nutrient application for wheat and maize yields and use efficiency of fertilizers in wheat/maize rotation system in lime concretion black soil area of southeast Henan province or other similar ecological regions.

Regulatory Function of Trehalase Genes on Chitin Metabolism in the Cuticle of Nilaparvata lugens

ZHANG Lu, ZHU ShiCheng, ZHENG Hao, SHEN QiDa, WANG ShiGui, TANG Bin

Scientia Agricultura Sinica. 2017, 50(6): 1047-1056. doi:10.3864/j.issn.0578-1752.2017.06.006

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【Objective】The previous research results showed that insect trehalase (TRE) can regulate chitin metabolism and control the molting process. In this study, the brown planthopper (Nilaparvata lugens) molting process, the changes of the expression of chitin content and chitin synthase (CHS) and chtinase (Cht) genes were detected when TRE genes were knocked down by the way of RNAi, in order to explore the roles of different trehalase genes in the regulation of chitin metabolism in the epidermis.【Method】N. lugens fed in the lab was chosen as the experimental material, and RNAi technology was used to inhibit the single or two TRE genes’ expression by injection of double stranded RNA. The total RNA was extracted from the cuticle of N. lugens using the TRIzol^®reagent as instructed by manufacturer. First-stand cDNA was synthesized using the PrimeScript^TM RT reagent Kit with gDNA Eraser following the manufacturer’s instructions. And the effect of RNAi was firstly determined after 48 h of injection by quantitative real-time PCR (qRT-PCR). Secondly, the chitin content of N. lugens whole body was determined at 48 h using potassium hydroxide method qRT-PCR, and photos of the insects with molting difficulties were taken in the same time. In the last, the relative expression levels of CHS and Cht of N. lugens were detected by qRT-PCR, and the regulatory function on chitin metabolism of TRE was analyzed at the same time. 【Result】Compared with the injection of dsGFP which was used as a control group, the results showed that in other groups injected with dsRNA the chitin content of N. lugens was decreased significantly, in which the dsTRE1 mixed injection group and the Validamycin injection group showed a significant decrease, meanwhile its molting problems appeared at the same time. qRT-PCR results showed that the gene expression of individual TRE was inhibited at 48 h after one TRE dsRNA injection, and the other TRE expression was increased and indicated it has a complementary function. The expressions of TRE1-2 and TRE2 were decreased in all groups, and dsTRE1s also inhibited the expression of TRE2. Secondly, the obvious effects could be found when mixed dsTRE1 trehalase inhibitor Validamycin injected into N. lugens and TRE genes were decreased significantly. The expression level of CHS and its splicing variants had no obvious effect when every TRE genes’ expression was knocked down, while CHS1 and CHS1a expressions were significantly decreased at 48 h after Validamycin injection. The expression of CHS1 in the cuticle increased after dsTRE1-2 injection and the expression of CHS1a increased after injection of dsTRE1-2. Thirdly, the expression levels of Cht1 and Cht8 decreased or decreased significantly after four dsTRE and Validamycin injection. The expression levels of Cht2 and Cht5 increased significantly when dsTRE1 was injected, as well as Cht2 and Cht4 increased significantly while Cht1, Cht6 and Cht8 decreased after dsTRE1-2 injection and Cht2 expressed increased significantly while the expression of Cht1, Cht8 and Cht10 decreased at 48 h when TRE2 knocked down. In the same time, the expressions of Cht1 and Cht5 decreased significantly while Cht9 increased significantly at 48 h after dsTRE1s injection. In the last, about all of 10 chitinase genes’ expression decreased significantly or extremely significantly after Validamycin injection.【Conclusion】TRE can control the synthesis of chitin through the regulation of chitin metabolic pathways in N. lugens. The results of this study will provide a theoretical basis for developing and screening effective trehalase inhibitors to control N. lugens.

Eukaryotic Expression, Affinity Purification and Enzyme Activity of Chitin Deacetylase in Locusta migratoria

ZHAO Pan, ZHANG XueYao, LIU XiaoJian, ZHAO XiaoMing, YU RongRong, DONG Wei, MA EnBo, ZHANG JianZhen, ZHANG Min

Scientia Agricultura Sinica. 2017, 50(6): 1057-1066. doi:10.3864/j.issn.0578-1752.2017.06.007

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【Objective】 The objective of this study is to investigate the eukaryotic expression and enzyme activity of chitin deacetylase 1 and 2 (LmCDA1 and LmCDA2) in Locusta migratoria. The results will provide an experimental basis to further clarify the physiological function of LmCDA1 and LmCDA2 in chitin degradation pathway and the development of new green pesticides.【Method】The domains of LmCDA1, LmCDA2a and LmCDA2b were predicted using BLASTP and SMART softwares. The full-length sequences of LmCDA1, LmCDA2a and LmCDA2b were obtained by PCR. Recombinant plasmids of pFastBac-LmCDAs were constructed, the recombinant Bacmid plasmids were obtained by transformation, and then transfected into Sf9 insect cells to express target proteins in vitro. Target proteins were detected by Western blot technology, and then purified by Ni-NTA affinity chromatography column and anion exchange chromatography column (Q-Sepharose). The purity of proteins was detected by 12% SDS-PAGE, and the enzymes activity was detected by spectrophotometric method with p-nitroacetylaniline as substrate. The method of T test was used to analyze the difference of enzyme activity of LmCDA2a and LmCDA2b. 【Result】 The results by using BLASTP and SMART softwares showed LmCDA1, LmCDA2a and LmCDA2b contained four structural domains, that are signal peptide, chitin binding peritrophin-A (ChBD), low-density lipoprotein receptor class A (LDLa) and catalytic domain (CDA). Three genes contained six conserved cysteines in ChBD. The spacing and amino acid composition between cysteines 3 and 4 (67-84 aa) and the sequence between cysteines 4 and 6 (84-106 aa) differed between LmCDA2a and LmCDA2b, the rest were identical. Western blot result showed that the protein molecular weight of LmCDA1, LmCDA2a and LmCDA2b was about 61 kD, consistent with the prediction, which suggested that the recombinant Bacmid plasmids were expressed successfully in Sf9 insect cells. The purity of purified proteins was detected by 12% SDS-PAGE electrophoresis. Most impurities could be removed by Ni-NTA affinity chromatography, and the proteins were further purified by Q-Sepharose exchange chromatography. Enzyme activity determination showed that LmCDA1, LmCDA2a and LmCDA2b had chitin deacetylase activity of 0.268, 0.354 and 0.228 U·μL^-1, respectively, and the activity of LmCDA2a and LmCDA2b showed significant differences. 【Conclusion】Eukaryotic expression in vitro and enzyme activity determination of LmCDA1, LmCDA2a and LmCDA2b revealed that these three enzymes have chitin deacetylase activity, and LmCDA2a and LmCDA2b enzyme activity has a significant difference. It was speculated that the reasons of different phenotypes of L. migratoria when LmCDA2a and LmCDA2b were silenced might be that LmCDA2a and LmCDA2b enzyme activity had a significant difference.

Capacity and Characteristics of Soil Microbial Biomass Under Various Climate and Fertilization Conditions Across China Croplands

WANG ChuanJie, XIAO Jing, CAI AnDong, ZHANG WenJu, XU MingGang

Scientia Agricultura Sinica. 2017, 50(6): 1067-1075. doi:10.3864/j.issn.0578-1752.2017.06.008

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【Objective】Soil microbial biomass is an important index of soil biological fertility and also is an important pool of soil nutrient turnover. The objective of this study was to explore the capacity and characteristics of soil microbial biomass carbon (SMBC) and nitrogen (SMBN) under various climatic zones and fertilization managements across China cropland, and further to obtain better understanding of the factors and how to manage soil biological fertility.【Method】The publications were collected by searching Web of China Knowledge Resource Integrated Database, Wan Fang Database and Web of Science with the keywords of "soil microbial biomass", "Chinese cropland" and "long-term fertilization". A data set with 458 pairs of reported soil organic carbon (SOC) and SMBC and 414 pairs of reported total nitrogen (TN) and SMBN was set up from 42 published papers. They included such four categories of climatic zone as typical temperate (M1: humid; M2: semi-arid), warm-temperate (W) and subtropical (S) zones and two categories of fertilization with (+OM) and without (-OM) organic amendments across China. Statistical analysis on the median of SMBC/SOC and SMBN/TN under different fertilization managements and climate zones was performed with Kruskal-Wallis H test (P<0.05). The capacity of SMBC and SMBN to SOC and TN was obtained from boundary line, respectively.【Result】Results showed that there was a significant response of soil microbial biomass (SMBC and SMBN) to the increase in SOC and TN under different fertilization managements and climate zones (P<0.01), respectively. The response coefficients (i.e., the correlation slope) between SMBC and SOC, SMBN and TN were 24.77 and 30.27 under organic amendments, respectively, which were significantly higher than that under the management without organic amendments (19.88 for SMBC and 19.86 for SMBN) (P<0.05). Boundary analysis showed that the maximum response coefficients of SMBC to SOC and SMBN to TN were 33.45-36.00 and 45.45-49.79, respectively. Compared with the maximum values, the current average values still have 37.99% and 49.66% of room for improvement, respectively. The median values of SMBC/SOC and SMBN/TN were significantly different among the four climate zones. The highest value of SMBC/SOC was 2.73% under temperate semi-arid and sub-humid zone (M2), followed by sub-tropical (S, 2.45%) and warm humid zone (W, 2.31%), the lowest value is 1.48% under temperate humid zone (M1). The sequence of SMBN/TN was W (4.72%) >M2 (3.50%) >S (2.99%) >M1 (1.80%). The ranges of SMBC/SOC and SMBN/TN were 0.35%-6.50% and 0.50%-9.72%, respectively, under various fertilization conditions with no significant differences. For different fertilization managements under similar climatic zone, the median value of SMBC/SOC and SMBN/TN was significantly higher under organic fertilizer management than that under no organic fertilizer management under temperate humid zone (P<0.05).【Conclusion】Climatic conditions have a significant impact on soil microbial biomass capacity. Different fertilization managements do not significantly change the SMBC/SOC and SMBN/TN, but the organic amendment can significantly improve SMBC and SMBN, which are helpful for soil biological fertility improvement.

Effects of Reduced N Fertilization on Crop N Uptake, Soil Ammonia Oxidation and Denitrification Bacteria Diversity in Maize/Soybean Relay Strip Intercropping System

ZHOU Li, FU ZhiDan, DU Qing, CHEN Ping, YANG WenYu, YONG TaiWen

Scientia Agricultura Sinica. 2017, 50(6): 1076-1087. doi:10.3864/j.issn.0578-1752.2017.06.009

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【Objective】The aim of this study is to reveal the variation characteristics of soil bacteria quantity and community in crop rhizosphere, and the relationship among bacteria quantity and community, soil total N content, crops N uptake in maize/soybean relay strip intercropping system, which will provide theoretical and technology support to reduce fertilizer input and efficient production in cereal/legumes intercropping system.【Method】Field experiments were conducted with two factors split-plot designed from 2013 to 2015, the main factor was planting patterns which included monoculture maize (MM), solo soybean (SS) and maize/soybean relay strip intercropping system (IMS), and the sub-factor total N for maize and soybean were no nitrogen 0 (NN), reduced nitrogen 180 kg N·hm^-2 (RN), conventional nitrogen 240 kg N·hm^-2 (CN), respectively. At V12, VT, R6 stages of maize and V5, R2, R5, R8 stages of soybean, the rhizosphere bacteria quantity was investigated by dilution-plate method, non-rhizosphere soil and crops shoot total N contents were determined by Kjeldahl method, and the diversity and gene abundance of ammonia oxdizing bacteria (amoA) and denitrifying bacteria (nirS) under different N fertilization rates were measured by combining clone library with FQ-PCR. 【Result】 The bacteria quantity in the rhizosphere of intercropped maize (IM) was 2.6% higher, and that of intercropped soybean (IS) was 12.9% higher compared with the corresponding monoculture. The total soil and shoot N contents of intercropped maize were increased by 13.39% and 2.10%, and that of intercropped soybean were decreased by 5.81% and 3.24% in contrast with corresponding monoculture, respectively. The gene abundance of ammonia oxdizing bacteria (amoA) of intercropped maize and soybean was 38.5% and 64.8% higher, and the gene abundance of denitrifying bacteria (nirS) of intercropped maize and soybean was 57.77% and 126.39% higher in contrast with corresponding monoculture, respectively. Under different N fertilization rates, the bacteria quantity in the rhizosphere of maize was 9.6% and 9.8% higher, and that of soybean was 11.7% and 11.0% greater in RN compared NN with CN, respectively. In addition, reduced N fertilization increased the diversity index of amoA gene in the rhizosphere of maize and soybean, and increased the diversity index of nirS of monoculture maize, moreover, reduced N fertilization decreased the diversity index of nirS of intercropped maize and soybean. 【Conclusion】 Reduced N fertilization contributed to increase bacteria quantity in the rhizosphere under maize/soybean relay strip intercropping system, improved the progress of soil N transfer, promoted the N uptake of maize and soybean thereby achieved saving fertilizer.

Effects of Shading Fruit with Opaque Paper Bag on Transcriptome in Peach

HE Ping, LI LinGuang, WANG HaiBo, CHANG YuanSheng, LI HuiFeng

Scientia Agricultura Sinica. 2017, 50(6): 1088-1097. doi:10.3864/j.issn.0578-1752.2017.06.010

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【Objective】 The objective of this study was to find the transcriptome differences between shading fruits with opaque paper bags and CK in peach, and enrich the peach transcriptome data. 【Method】 Selecting peach fruits as samples (shading fruits with opaque paper bags and CK, respectively), then the transcriptome libraries of peach fruits were constructed by using the Illumina HiSeqTM 2500 sequencing technique, and were analyzed by using the bioinformatics methods subsequently, such as sequencing assess and gene function annotation. 【Result】 The results showed that the 16.62 Gb transcriptome data were obtained from both the shading bagged fruits and CK , and the base ratios with quality values higher than 30 in reads (Q30) were more than 91% from both the shading fruits and CK. There were 65 300 730 reads and 66 603 686 reads transcriptome data were obtained from the peach fruits used bagging or not, and 85.73% and 84.60% reads could be compared to the peach reference genome sequence, respectively. Compared the transcriptome of shading bagged fruits with CK, 1 963 differentially expressed genes were found in the shading bagged fruits, including 1 255 up-regulated genes and 708 down-regulated genes. Annotation analysis indicated that 1 957 genes were annotated in Nr data, including 1 252 up-regulated genes and 705 down-regulated genes. Through COG analysis, there were 853 functional annotations of these differentially expressed genes, involving 23 functional classifications. With GO function annotation classifications, a total of 1 609 genes were divided into 53 function categories, in which many functional categories were mainly involved, such as molecular binding, catalytic activity, cell process, biological regulation. KEGG analysis showed that a total of 421 genes were annotated to 94 metabolic pathways, and photosynthesis signaling pathway, carbon fixation in photosynthetic organisms, photosynthesis-antenna proteins signaling pathway, flavonoid biosynthesis, ribosome biosynthesis (DNA replication) pathway were significantly enriched. The photosynthesis and flavonoid biosynthesis signaling pathway played an important role in fruit color, but the role of ribosome biosynthesis was not clear. The fruit quality also was detected from the shading bagged fruits and CK. The results showed that shading bagged peach fruits had a remarkable impact on soluble solids and total soluble sugar, which were significantly reduced. But the study also found, on average, little or no effect on total acid and per fruit weight from both the shading fruits and CK. 【Conclusion】 The number of differentially expressed genes under different physiological status of the fruit tissue in peach were obtained. The study found that the photosynthesis and flavonoid biosynthesis signaling pathway played an important role in fruit color and shading bagged peach fruits had a remarkable impact on soluble solids and total soluble sugar.

Analysis of the Genetic Diversity of Wild Rosa roxburghii Populations in Guizhou Province Based on EST-SSR Marker

ZHANG HuaiShan, YAN XiuQin, LU Min, WANG DaoPing, AN HuaMing

Scientia Agricultura Sinica. 2017, 50(6): 1098-1108. doi:10.3864/j.issn.0578-1752.2017.06.011

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【Objective】Genetic structure and genetic diversity of wild Rosa roxburghii resources were analyzed and evaluated in order to provide a scientific basis of the protection and excavation of R. roxburghii resources.【Method】Ten EST-SSR primers with good amplification and high polymorphism were used to analyze the genetic diversity and genetic structure within 12 populations including 255 R. roxburghii germplasms by POPGENE 1.31 software. According to Nei’s standard genetic distance, NTSYS-pc2.10e software was used to cluster the populations, and the Mantel correlation relationship between Nei's genetic distance and geographical distance was tested.【Result】The range of Shannon’s information index (I) was from 0.62 to 0.88, Nei’s gene diversity from 0.40 to 0.52. Total percentage of polymorphic loci (P) was 100% in these populations, except for Fuquan and Qinglong populations. The Chi-square test results showed that the 12 populations of R. roxburghii did not follow with the Hardy-Weinberg Equilibrium at most loci, and the F_it and F_is values were negative. Characteristics, such as F_st =0.0403, gene flow N_em=5.9484, genetic identity (0.9068<GI<0.9926) and genetic distance (GD<0.0978) were discovered among the 12 populations. Futhermore, Mantel test revealed a significant correlation between genetic and geographic distance among populations (r=0.2498, P=0.9512).【Conclusion】The 10 pairs of EST-SSR primers are highly polymorphic genetic markers which can be used effectively for evaluating genetic structure and genetic diversity of R. roxburghii. R. roxburghii in Guizhou province has moderate to high diversity. The vast majority of genetic variation and an excess of heterozygosity occur within populations. And among the populations, it has the characteristics of frequent gene exchange, high genetic consistency and small Nei 's genetic distance.

Enantiomeric Analysis of Volatile Terpenoids in Different Teas

SHAO ChenYang, Lü HaiPeng, ZHU Yin, ZHANG Yue, LIN Zhi

Scientia Agricultura Sinica. 2017, 50(6): 1109-1125. doi:10.3864/j.issn.0578-1752.2017.06.012

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【Objective】 Volatile terpenoids are important components of tea aroma, and most of them contain chiral carbon structure which lead to the existence of corresponding enantiomers with different aroma characteristics. Identifying the terpenoid enantiomers among different teas can result in a better understanding of the mechanisms underlying tea aroma formation for tea quality improvement. 【Method】 An analytical approach for enantiomeric analysis of volatile terpenoids in tea was firstly established by headspace solid phase microextraction combined with chiral gas chromatography-mass spectrometry. The chiral capillary column of Supelco β-DEX110 was used for terpenoid enantiomer analyses with corresponding authentic standards. And the compositions of enantiomers of terpenoids in representative green teas, black teas, oolong teas, white teas and dark teas were then investigated. 【Result】 Nine pairs of terpenoids enantiomers were successfully separated. The data of experiment indicated that linalool and linalool oxide B enantiomers were detected in all tea samples with high contents: S-linalool was considered as the major configuration of linalool in most kinds of teas, except for Yingde and Yunnan black teas and Indonesia white tea, which contained R-(-)-linalool as the major configuration of linalool; linalool oxide B presented the same stereo configuration in green tea, black tea, white tea and dark tea, whereas in some oolong teas (Lingtou Dancong and Wenshan Baozhong) a higher proportion of opposite configuration existed. Linalool oxide A was detected in green and dark teas with the same major stereo configuration, while it presented reverse major configuration in some black teas (Keemun and Darjeeling black teas) and oolong teas (Shuixianwudong Dancong and Tieguanyin). The single and consistent configurations of linalool oxides C and D were observed in most tea samples. The other important terpenoids distributed in various situations among different tea samples: α-pinene was only detected in the Lapsang Souchong tea, and the major configuration was S. different configurations and various enantiomeric excess values of α-terpineol were observed in different teas. R-(-)-4-terpineol was the major configurations of 4-terpineol in all tea samples, and S-(+)-4-terpineol could only be detected in the Liu Pao tea (dark tea), and a single R-configuration of α-ionone could be detected in most teas (except green teas). The quantitative results indicated that the highest contents in tea aroma of most tea samples were contributed by linalool, and the ranking order of mean contents of linalool was white tea>black tea>oolong tea>green tea>dark tea. the contents of linalool oxide B frequently second to those of linalool, and the ranking order of mean contents was black tea>white tea>oolong tea>dark tea>green tea. the contents of other terpenoids were generally lower than 100 ng·g^-1, and the content distributions were widely different among various tea samples. 【Conclusion】 This study investigated the content variations of the nine pairs of important terpenoid enantiomers in different kinds of tea for the first time. The present research result could contribute to get a deeper understanding of the chemical substances of tea aroma, and provide a new thought for the quality control of tea aroma, the discrimination of tea cultivars and origins and quality certification of tea.

Effects of Eight Chinese Style Cuisine Methods on Polycyclic Aromatic Hydrocarbons, Trans-Fatty Acids and Nitrite of Beef

ZHANG Lan, GAO TianLi, LIU YongFeng, ZHAO Jing, LIAO Jing, KU Ting

Scientia Agricultura Sinica. 2017, 50(6): 1126-1138. doi:10.3864/j.issn.0578-1752.2017.06.013

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【Objective】The purpose of this study is to select the processing techniques of beef with less harmful substances from the steaming, boiling, braising, broiling, frying, pan-frying, drying and pickling eight traditional Chinese cooking process. The result of the present study will also provide a theoretical basis for consumers to choose a reasonable cuisine method with less harmful substances.【Method】By adjusting the temperature and time of broiling, frying and pan-frying, the contents of polycyclic aromatic hydrocarbon (PAHs), trans-oleic acids (C18: 1 trans-9) and nitrite are compared, so that the optimum cooking conditions of broiling, frying and pan-frying were selected. On this basis, the eight Chinese style culinary arts were comprehensively analyzed so as to select the optimum cooking methods, and the indexes were determined by high performance liquid chromatography (HPLC) external standard method, gas chromatography-mass spectrometry (GC-MS) external standard method and spectrophotometry, respectively.【Result】Four PAHs including pyrene, benzo[a] anthracene, chrysene and benzo [k] fluoranthene were detected in control group. The treatments of broiling at 160-180℃ were better, and the contents of the five PAHs were lower. The contents of PAHs were lower in process of frying for 3-4 min and pan-frying for 2-3 min. Among the eight Chinese style cuisine methods, the processes of frying and pan-frying had the highest number of PAHs species. The benzo [b] fluoranthene content in processed of frying and pickling were higher. The chrysene contents in process of steaming and pan-frying were higher, the benzo [b] fluoranthene contents in processes of broiling, frying and pan-frying were higher, the contents of benzo [k] fluoranthene, benzo[a] pyrene, Dibenz [a, h] anthracene in frying and pan-frying treatments were higher. The C_{18:1 trans-9} content in the process of frying for 3min and pan-frying for 2 min were lower (P＜0.05). The effect of broiling temperature on the content of C_{18:1 trans-9}was not significant. The C_{18:1 trans-9} contents in frying and pan-frying treatments were higher among the eight Chinese style culinary arts (P＜0.05). The nitrite content in the process of broiling at 160℃ and 180℃ were lower, and that in treatments of frying for 3 min and pan-frying for 2 min were the lowest (P＜0.05). The nitrite contents in treatments of frying, pan-frying and pickling were higher among the eight Chinese style culinary arts (P＜0.05).【Conclusion】The content of three harmful substances in beef that being broiled for 40 min at the temperature of 160℃, at the temperature of 226-228℃ fried for 3 min and pan-fried for 2 min were lower compared with the control group. A comprehensive analysis revealed that the beef processed by steaming, boiling, braising and drying contains less hazardous substances.

Effect of EDN3 on of Sheep Skin Melanocytes with Different Coat Colors in Vitro

LI YaNan, ZHAO BingLing, WANG HaiDong, CHEN TianZhi, LIU Ying, CHANG LuCheng, DONG ChangSheng

Scientia Agricultura Sinica. 2017, 50(6): 1139-1146. doi:10.3864/j.issn.0578-1752.2017.06.014

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【Objective】 The objective of this study is to explore the effects of EDN3 in vitro on sheep melanocytes with different coat colors and to find the mechanisms of production of melanin and their differences. 【Method】 Melanocytes from black and white coat colors were cultured in vitro, and MTT assay was used to detect different cell proliferation rates by affected EDN3. After extraction of total RNA and total protein of both cells, the total RNA was reversely transcribed to cDNA, and then the effect of EDN3 on mRNA relative expression of EDNRB, NRas and TYR was analyzed by RT-PCR, and Western blot was performed to ensure whether EDN3 had influence on the protein expression of EDNRB, NRas and TYR. Significant difference was analyzed by using statistical method with software SPSS19.0. 【Result】 MTT assay showed that EDN3 promoted cells proliferation from both black and white coat colors. RT-PCR revealed that compared with the control group, the relative expressions of EDNRB and NRas mRMA in EDN3 group from white coat color were significantly increased by 1.7992 folds and 1.8536 folds respectively (P<0.01), while TRY mRNA had no significant difference. In cells from black coat color, the expressions of EDNRB, NRas and TYR were all induced significantly with multiples of 2.2512, 1.3859 and 15.5710 respectively (P<0.01). The result of Western blot in EDN3 group was consistent with RT-PCR, the EDNRB protein expression from white coat color was 3.0827 times of control group (P<0.01), and the NRas protein expression was 1.2936 fold of control group (P<0.05), while TYR had no change. The protein expressions in cells from black coat color of EDNRB, NRas and TYR were observably added up to 3.9800 time (P<0.01), 1.3658 and 1.8498 times (P<0.05), respectively. 【Conclusion】 EDN3 promoted melanocytes proliferation from both black and white coat colors, while had no effect on TYR which is a rate-limiting enzyme of pigment synthesis in cells from white coat colors, and might play a role in producing melanin in cells isolated from black coat color.

Time Course Effect of Lipopolysaccharide on Toll-Like Receptors Expression in the Goose Follicular Stroma

YING ShiJia, DAI ZiChun, GUO JiaJia, SHI ZhenDan

Scientia Agricultura Sinica. 2017, 50(6): 1147-1156. doi:10.3864/j.issn.0578-1752.2017.06.015

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【Objective】 This study aims to investigate the transcriptional profiling of Toll-like receptors (TLRs) and their responses to lipopolysaccharide with different treatment times in the goose follicular stroma. 【Method】 The laying process was monitored in a flock of Yangzhou geese. The geese were injected intravenously with LPS (1.5mg/kg body weight) 8 and 2 h after oviposition, and 4, 16 and 28 h before oviposition. All experimental geese were slaughtered 8 h after oviposition. Therefore, time course of LPS was achieved, namely at 0, 6, 12, 24 and 36 h after injection of LPS. Five geese at each time point were selected. After slaughtering, the ovaries were collected, the follicle morphology was observed, and the stroma of the first largest follicles (F1), F2, F3, F4 and F5 was isolated. The animals were provided with feed and water ad libitum, and maintained under natural photoperiod. For the geese 0, 24 and 36 h of after LPS treatment, all samples were used for gene expression analysis. For the other geese, RNA samples of follicular stroma of each goose were mixed with equal concentrations for gene expression analysis. RT-PCR was performed to examine the transcriptional profiling of 10 types of avian TLRs in follicular stroma. The spleen tissue was used as the positive control, and the sample without cDNA sample was used as the negative control. The expression levels of TLRs in follicular stroma among different hierarchical follicles and among different time points were tested using Real-time PCR. For data on the time course of LPS and different hierarchical follicles, the significance of differences was analyzed using the one-way ANOVA followed by Duncan’s multiple range tests. For data between stroma in control group and DF, statistical analysis was carried out using independent-samples T test. 【Result】 All 10 reported TLRs in poultry, namely TLRs 1A, 1B, 2A, 2B, 3, 4, 5, 7, 15 and 21 were expressed in goose stroma of hierarchical follicles. The expression level of TLR2A exhibited a tendency to increase with follicular growth. The TLR2A expression in F1 was higher than in F3, F4 and F5, and the TLR15 expression in F1 was higher than in F5. There were no significant effects of follicle sizes on the expression of TLRs 1A, 1B, 2B, 3, 4, 5, 7 and 21 in stroma. The morphology and colour of ovarian follicles were not changed at 0, 6 and 12 h after administration of LPS. However, the hierarchical follicles of three birds after 24 h and all birds after 36 h became an irregular ellipse or circle in shape and deep yellow in colour. Compared with the control (LPS treatment 0 h), the expression of TLRs 2A, 4 and 5 was significantly increased at 12 and 24 h after LPS treatment, the expression of TLR2B was significantly increased at 24 h, and the expression of TLRs 7 and 15 was significantly increased during the 6 to 24 h period. LPS stimulation did not significantly affect the expression of TLRs 1A, 1B, 3 and 21 during the 6 to 24 h period. Compared with the control, the expression of TLRs 1A, 2A, 2B, 4, 5, 7 and 15 was significantly increased in the denatured hierarchical follicles at 24 and 36 h, while the expression of TLRs 1B and 21 was significantly increased in the denatured hierarchical follicles at 36 h. 【Conclusion】 All the 10 members of avian TLR families are expressed in goose follicular stroma. Furthermore, with prolonged LPS treatment, the morphology of hierarchical follicles is changed, but the TLRs expression levels are still increased.

De novo Transcriptome Assembly for Apis cerana cerana Larval Gut and Identification of SSR Molecular Markers

XU XiJian, GUO Rui, LUO Qun, XIONG CuiLing, LIANG Qin, ZHANG ChuanLian, ZHENG YanZhen, ZHANG ZhaoNan, HUANG ZhiJian, ZHANG Lu, LI WenDong, CHEN DaFu

Scientia Agricultura Sinica. 2017, 50(6): 1157-1166. doi:10.3864/j.issn.0578-1752.2017.06.016

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【Objective】 The objective of this study is to de novo assemble a reference transcriptome for Apis cerana cerana larval gut, perform gene function and pathway annotation for this transcriptome, and to identify specific SSR molecular markers for A. c. cerana larvae. 【Method】 3-day-old instar A. c. cerana larvae were fed with the purified Ascosphaera apis spores, the guts of 4-, 5- or 6-day-old honeybee larvae were sampled and used as sequencing material for RNA seq. After filtration, clean reads were obtained, and unigenes were assembled using Trinity software. BLASTX tool (E-value＜10⁻⁵) was used to search the unigenes against NCBI Nr, Swiss-Prot protein, KOG as well as KEGG databases to perform gene function and pathway annotation. MISA software was used to search microsatellite markers in the larval gut’s transcriptome. The specific primers of all SSRs were designed using Primer Premier 5 program and several pairs were used to amplify SSR loci in A. c. cerana larvae samples from 3 different regions (Beijing, Xingcheng, and Chengdu) in China by method of PCR. 【Result】 In this study, RNA seq produced 35 670 000 high quality reads, which were assembled into 43 557 unigenes with a mean length of 898 nt. 18 225 unigenes were annotated in the public protein databases. a total of 11 442 unigenes had a KOG classification and they distributed in 25 KOG categories, among them, RNA processing and modification was the largest group (1 249). 9 679 unigenes could be classified into three gene ontology (GO) categories, in which the mostly enriched ones were cellular process (4 201 unigenes), cell (2 900 unigenes) and binding (4 935 unigenes). 4 517 unigenes were annotated to 216 KEGG pathways, among them, ribosome (385 unigenes) was the largest. Finally, 13 448 SSRs were found in 7 763 unigenes, and 6 out 20 SSR loci could be successfully amplified in A. c. cerana larvae samples from 3 different regions in China using PCR. 【Conclusion】 This study assembled and annotated a reference transcriptome for A. c. cerana larval gut, which will provide a key information not only to studies on eastern honeybee and its larvae such as molecular biology and omics, but also to improve and validate the genome of A. cerana. SSR markers developed here could be applied to future investigation of A. c. cerana including gene map construction, genetic diversity analysis as well as gene location. Meanwhile, this study suggested that developing molecular markers using transcriptome data of non-model organism is a rapid and efficient method.

Transcriptome Difference Analysis of Saccharum spontaneum Roots in Response to Drought Stress

LIU HongBo, LIU XinLong, SU HuoSheng, LU Xin, XU ChaoHua, MAO Jun, LIN XiuQin, LI ChunJia, LI XuJuan, ZI QiuYan

Scientia Agricultura Sinica. 2017, 50(6): 1167-1178. doi:10.3864/j.issn.0578-1752.2017.06.017

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【Objective】The objective of this study is to understand the inner molecular mechanisms of Saccharum spontaneumclone named Yunnan 82-114 in response to water stress, and mine these genes closely related to drought tolerance, improve the utilization efficiency of Saccharum spontaneum in sugarcane drought-resistant breeding program. 【Method】llumina HiSeqTM 4000, a high-through transcriptome sequencing technology, was applied to obtain the transcriptome differential expression data of Yunnan82-114 roots under drought stress treatment for 24h, 48h, and 72h. These assembled unigenes above were compared with database of Swiss-Prot, Nr, KOG, Pfam, and KEGG, respectively, then the abundance of gene expression among different samples were screened according to transcriptome data by using RPKM method, and the differentially expressed genes among the treated samples were estimated by referring to the standard of FDR≤0.05 & |log2 fold change|≥1. Function and pathway of those different expression genes were also investigated using Gene Ontology（GO）database and KEGG pathway database.【Result】Total 134 724, 130 368, 133 564, and 131 321 expressing genes were got, respectively, from the untreated control and three treated samples(24 h, 48 h and 72 h). Compared the control with the treated samples, about 3 061 (1 302), 2 304 (2 841) and 3 236 (2525) genes whose expression was significantly up-regulated (or down-regulated) were detected, respectively. When the FDR value was set to be 0 as a selection criterion, some genes performing extreme significantly different expression between control and treated samples were acquired, they mainly involved in transcriptional activation, water transport, DNA binding, ATP binding, membrane/ transmembrane transport, and defense response. GO enrichment analysis showed that some differences in the biological approach categories existed among the treated samples, for the samples treated for 24 h and 48h, the most enriched category was DNA-dependent transcription and transcription factor, then to the samples treated for 72 h, the enriched category was the translation-related genes. At the aspects of cellular components and molecular functions, the enriched category was inner membrane, nucleus, and ATP binding related genes in the three treated samples. KEGG enrichment analysis illustrated that 2 248 differentially expressed genes involved in 107 pathways in 24 h-treated sample, 2 114 in 130 in 48h-treated sample, and 2 392 in 144 in 72 h-treated sample. Finally, some genes related to abiotic stress and resistance were screened via KEGG significance enrichment analysis, which include glycosylsphingolipid biosynthesis, MAP kinase signal transduction, and ABC translocator.【Conclusion】Expression pattern and function information of differentially expressed genes between three water-stressed samples and a control were analyzed. Some genes like extracellular signal-regulated kinase, phospholipase D, hexosaminidase, and ATP-binding cassette B-1 were found to involve in response to water stress for Yunnan82-114. About 70 significant up-regulated expression genes at transcription level were obtained, but there were 11 genes exhibiting down-regulated expression, this implied that these genes induced or suppressed fiercely by drought stress have obvious relationships with drought tolerance of Yunnan82-114.

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