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Crop Science

Ethanol production and mitochondrial-related gene expression of maize (Zea mays) seed during storage 

✺ Chalisa Chaengsakul, Damrongvudhi Onwimol, Pasajee Kongsil,Sawita Suwannarat

Mitochondrial degradation plays a vital role in seed deterioration. Novel markers were investigated based on a new method for quantifying maize seed deterioration during 12 months' storage under ambient (lab bench, ~27°C and 50–80% relative humidity (RH)) or controlled ((15±1)°C and (50±5)% RH in bags with low oxygen permeability) conditions involving two techniques: 1) fast ethanol assay and 2) quantitative RT-PCR (qPCR) with four mitochondrial-related genes in maize seed: alcohol dehydrogenase (ADH1), alternative oxidase (AOX1), cytochrome c oxidase (COXc), and ATPase. Ethanol production during imbibition and the expression of genes using the new method were compared to the results of two conventional methods: a germination test and an accelerated ageing test. The results showed that germination following ambient seed storage reduced significantly compared to the controlled conditions, especially at 9 months of storage. Ethanol production of maize seed measured by fast ethanol assay increased during storage. After 6 months, the mean (n=4) ethanol production from seed under ambient conditions was 400 µg L^–1 which was higher than under the controlled conditions (240 µg L^–1). Stored mRNA level of COXc and ATPase significantly decreased over time in ambient storage but were quite stable under the controlled conditions. Maize seed was also treated for artificial ageing at 42°C in 100% RH for 12, 24, and 48 h. At 24 h after treatment (HAT), maize seed produced significantly more headspace ethanol than at 12 HAT and more than the control (non-treated seed). The transcription level of ADH1 and ethanol production increased. The transcription level of COXc was directly related to the severity of the ageing treatment. In conclusion, a combination of fast ethanol assay and qPCR enhanced understanding of maize seed deterioration and provided new possibilities for the evaluation of seed storability based on transcriptional levels. ↑ 向上滑动查看更多

Cite this article:   

Chalisa Chaengsakul, Damrongvudhi Onwimol, Pasajee Kongsil,Sawita Suwannarat. 2019. Ethanol production and mitochondrial-related gene expression of maize (Zea mays) seed during storage. Journal of Integrative Agriculture, 18(11): 2435-2445.

Genome-wide association study for starch content and constitution in sorghum (Sorghum bicolor (L.) Moench)

高粱籽粒淀粉及其组分的全基因组关联分析

✺ CHEN Bing-ru, WANG Chun-yu, WANG Ping, ZHU Zhen-xing, XU Ning, SHI Gui-shan, YU Miao, WANG Nai, LI Ji-hong, HOU Jia-ming, LI Shu-jie, ZHOU Yu-fei, GAO Shi-jie, LU Xiao-chun, HUANG Rui  

淀粉是高粱籽粒胚乳中最重要的成份之一。一般来说，淀粉以直链淀粉和支链淀粉两种形式存在胚乳中。直链淀粉和支链淀粉的含量对高粱籽粒的外观、结构和品质以及市场用途起着重要的作用。在本研究中，选取了634份高粱种质资源，其中包括242份来自国际半干旱研究所的微核心种质，252份我国地方品种和140份栽培种。该群体的平均淀粉含量为67.64%，平均直链淀粉和支链淀粉含量分别为20.19%和79.81%。同时，634份高粱种质资源进行了SLAF测序，获得了260,000高密度SNP标记。利用SLAF测序数据对该群体的淀粉含量、直链淀粉含量和直/支比例3个性状进行了基于混合性状模型的全基因组关联分析。共检测出70个与淀粉、直链淀粉含量和直/支比例有关的显著SNPs。在显著SNP两侧200kb的区间，通过GO分析筛选出与淀粉代谢有关的5个QTL12个候选基因，分别位于第1、5、6、和9染色体上。在12个候选基因中，基因Sobic.006G036500.1编码着肽基脯氨酰顺反异构酶 CYP38，它参与磷酸戊糖途径。基因Sobic.009G071800编码着6-磷酸-果糖激酶，是糖酵解途径中关键的限速酶。针对找到的候选基因，我们设计了KASP标记对50个淀粉含量高的种质和50个淀粉含量低的种质进行了基因分型，分型结果发现等位基因C与淀粉含量高相关、等位基因T与淀粉含量低相关，为高粱品质性状改良提供了可靠的单倍型数据。

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CHEN Bing-ru, WANG Chun-yu, WANG Ping, ZHU Zhen-xing, XU Ning, SHI Gui-shan, YU Miao, WANG Nai, LI Ji-hong, HOU Jia-ming, LI Shu-jie, ZHOU Yu-fei, GAO Shi-jie, LU Xiao-chun, HUANG Rui. 2019. Genome-wide association study for starch content and constitution in sorghum (Sorghum bicolor (L.) Moench). Journal of Integrative Agriculture, 18(11): 2446-2456. 

Transcriptome profiling using RNA-seq to provide insights into foxtail millet seedling tolerance to short-term water deficit stress induced by PEG-6000 

基于RNA-seq对不同谷子品种在PEG-6000模拟短期干旱胁迫下的转录组分析

✺ XU Bing-qin, GAO Xiao-li, GAO Jin-feng, LI Jing, YANG Pu, FENG Bai-li

谷子广泛种植在我国的干旱和半干旱地区，具有较高的抗旱性，是一种新的遗传和分子研究模式植物。为了揭示不同基因型谷子的抗旱性分子机制，本研究对两种谷子基因型'大毛毛谷'(DM)和'红粘谷'(HN)进行了抗旱生理和转录组综合分析。研究表明，在干旱条件下，基因型'DM'的表现优于'HN'，具体表现在DM叶片相对含水量RWC和叶绿素含量下降更为缓和。进一步的生理学和转录组学综合分析表明，这两种谷子基因型在干旱条件下，其细胞参与的一些重要的抗逆生物学途径具有高度的保守性，包括激素合成，脯氨酸和可溶性糖合成，以及活性氧(ROS)代谢等途径。但是，干旱条件下，两品种中与这些途径相关的部分基因则显示出了不同的表达模式。另外，HN中的MDA含量低于DM，并且干旱胁迫过程中无明显变化，原因可能是干旱条件诱导了HN在抗坏血酸 - 谷胱甘肽循环代谢途径中的更多的活化基因。总之，ABA代谢及信号传导相关基因，参与ROS抗氧化系统相关基因，参与脯氨酸和可溶性糖合成的相关酶基因，及通道蛋白基因和转录因子，共同组成了谷子响应干旱胁迫的早期策略。本研究对了解不同基因型的谷子响应短期干旱胁迫的生理及分子机制提供了基础，对于谷子抗旱品种选育具有重要意义。

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Cite this article:   

XU Bing-qin, GAO Xiao-li, GAO Jin-feng, LI Jing, YANG Pu, FENG Bai-li. 2019. Transcriptome profiling using RNA-seq to provide insights into foxtail millet seedling tolerance to short-term water deficit stress induced by PEG-6000. Journal of Integrative Agriculture, 18(11): 2457-2471.

The effects of soil moisture and salinity as functions of groundwater depth on wheat growth and yield in coastal saline soils 

滨海盐土水埋深对土壤水盐及小麦生长和产量的影响

✺ ZHANG He, LI Yan1, MENG Ya-li, CAO Nan, LI Duan-sheng, ZHOU Zhi-guo, CHEN Bing-lin, DOU Fu-gen 

水埋深会影响滨海盐土水分和盐分的含量，从而对作物生长和产量产生影响。本研究旨在探讨：1) 受水埋深影响的水盐胁迫对小麦生长的影响，2) 确定滨海盐土小麦生长较适宜的水埋深。大田试验水埋深处理 (通过土柱来维持) 分别在2013-2014年为0.7、1.1、1.5、1.9、2.3、2.7 m，2014-2015年为0.6、1.0、1.4、1.8、2.2、2.6 m。结果表明，土壤水分与盐分呈正相关关系。渍水与高盐胁迫 (2013-2014年水埋深0.7 m，2014-2015年0.6 m) 对小麦生长的影响程度大于轻度干旱、中盐胁迫 (2013-2014年2.3 m) 或轻度干旱、低盐胁迫 (2013-2014年2.7 m，2014-2015年2.2 m和2.6 m)。小麦孕穗期对水盐胁迫反应最敏感。在2013-2014年，水埋深 (沟深) 1.9 m (正常水分、中盐) 处和2.3 m (轻度干旱、中盐) 处，2014-2015年，水埋深1.8 m (正常水分、中盐) 处、2.2 m (轻度干旱、低盐) 处，小麦的叶面积、SPAD、净光合速率、籽粒产量和面粉品质最佳。在滨海盐土0-20 cm和20-40 cm土层下，较适宜的土壤相对含水量和土水比1:5浸提液电导率分别为58.67%-63.07%和65.51%-72.66% (2013-2014年)，63.09%-66.70%和69.75%-74.72% (2014-2015年)；0.86-1.01 ds m^-1和0.63-0.77 ds m^-1 (2013-2014年)，0.57-0.93 ds m^-1和0.40-0.63 ds m^-1 (2014-2015年)。

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ZHANG He, LI Yan1, MENG Ya-li, CAO Nan, LI Duan-sheng, ZHOU Zhi-guo, CHEN Bing-lin, DOU Fu-gen. 2019. The effects of soil moisture and salinity as functions of groundwater depth on wheat growth and yield in coastal saline soils. Journal of Integrative Agriculture, 18(11): 2472-2482.

Early selection of bread wheat genotypes using morphological and photosynthetic attributes conferring drought tolerance

✺ Hafiz Ghulam Muhu-Din Ahmed, Abdus Salam khan, LI Ming-ju, Sultan Habibullah Khan, Muhammad Kashif 

Genetic diversity is the base of any genetic improvement breeding program aimed at stress breeding. The variability among breeding materials is of primary importance in the achievements of a good crop production. Herein, 105 wheat genotypes were screened against drought stress using factorial completely randomized design at seedling stage to determine the genetic diversity and traits association conferring drought tolerance. Analysis of variances revealed that all the studied parameters differed significantly among all genotypes, indicating the significance genetic variability existed among all genotypes for studied indices. The 10 best performance genotypes G1, G6, G11, G16, G21, G26, G39, G44, G51, and G61 were screened as drought tolerant, while five lowest performance genotypes G3, G77, G91, G98, and G105 were screened as drought susceptible. Root length, chlorophyll a, chlorophyll b, and carotenoid contents were significantly correlated among themselves which exhibited the importance of these indices for rainfed areas in future wheat breeding scheme. Shoot length exhibited non-significant and negative association with other studied traits, and its selection seems not to be a promising criteria for this germplasm for drought stress. Best performance genotypes under drought stress conditions will be useful in future wheat breeding program and early selection will be effective for developing high yielding and drought tolerant wheat varieties.↑ 向上滑动查看更多

Cite this article:   

Hafiz Ghulam Muhu-Din Ahmed, Abdus Salam khan, LI Ming-ju, Sultan Habibullah Khan, Muhammad Kashif. 2019. Early selection of bread wheat genotypes using morphological and photosynthetic attributes conferring drought tolerance. Journal of Integrative Agriculture, 18(11): 2483-2491.

Spatial distribution of bolls affects yield formation in different genotypes of Bt cotton varieties

不同基因型抗虫棉品种棉铃空间分布对产量形成的影响

✺ NIE Jun-jun, YUAN Yan-chao, QIN Du-lin, LIU Yan-hui, WANG Shuang-lei, LI Jin-pu, ZHANG Mei-ling, ZHAO Na, GUO Wen-jun, QI Jie, MAO Li-li, SONG Xian-liang, SUN Xue-zhen

为优化棉铃空间分布结构，提高棉花产量，本试验选用三个产量构成差异较大的抗虫棉品种(岱字棉99B、鲁棉研21号和冀棉169)为试验材料，2013-2017年大田条件下，通过研究品种间农艺性状、棉铃空间分布特性，从而阐释不同基因型抗虫棉品种产量与纤维品质形成差异的原因。结果表明，冀棉169皮棉产量最高(5年平均42.2 g plant^-1)，分别比鲁棉研21号和岱字棉99B高16.3%和36.9%。产量构成因素中，只有单铃子棉重的变化趋势与产量基本一致，因此皮棉产量的提高取决于单铃子棉重的增加。然而，三个品种纤维品质的差异与皮棉产量不同，鲁棉研21号显著高于冀棉169和岱字棉99B，这主要归因于棉铃空间分布的差异。与岱字棉99B相比，株型松散的冀棉169单铃子棉重最大，内围结铃数最多；而株型紧凑的鲁棉研21号单铃子棉重较大，基部和中部果枝结铃数最多。在五年试验中，棉株内围铃对皮棉产量的贡献率，冀棉169、鲁棉研21号和岱字棉99B分别为80.72%、77.44%和66.73%。产量形成虽受年际变化的显着影响，但冀棉169的皮棉产量始终最高且最稳定，其产量构成各因素亦是如此。综上，棉花产量的提高主要归因于单铃重的增加，单铃重的增加和纤维品质的提高主要取决于棉铃空间分布结构的优化。

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NIE Jun-jun, YUAN Yan-chao, QIN Du-lin, LIU Yan-hui, WANG Shuang-lei, LI Jin-pu, ZHANG Mei-ling, ZHAO Na, GUO Wen-jun, QI Jie, MAO Li-li, SONG Xian-liang, SUN Xue-zhen. 2019. Spatial distribution of bolls affects yield formation in different genotypes of Bt cotton varieties. Journal of Integrative Agriculture, 18(11): 2492-2504.

Horticulture

Molecular characterization, expression and function analysis of eukaryotic translation initiation factor (eIF1A) in Mangifera indica

芒果翻译起始因子eIF1A基因克隆、表达与功能分析

✺ LI Li-shu, LUO Cong, AN Zhen-yu, LIU Zhao-liang, DONG Long, YU Hai-xia, HE Xin-hua

翻译起始因子eIF1A是蛋白合成重要的调控因子之一，在植物响应非生物胁迫过程中发挥重要的调控作用。但在芒果的果实成长发育和非生物胁迫中研究较少。本研究从芒果中克隆获得了1个MieIF1A-b基因，包含1个435 bp开放阅读框(ORF)，编码144个氨基酸(GenBank登录号：KP676599)，分子量为16.39 kDa，等电点为4.6；系统进化分析发现其与巨桉、木薯和毛果杨的同源性较高，分别为96%和95%；荧光定量表达分析表示MieIF1A-b在芒果的各组织中均有表达，尤其在80 d的成熟果实中表达量最高，在盐和干旱胁迫条件下均能诱导其在芒果叶片中的上调表达；转基因功能验证表明过表达MieIF1A-b基因能增强了转基因拟南芥的生长、表型和耐盐性。结果表明，MieIF1A-b基因与芒果的果实发育和盐害调控密切相关，为芒果非生物胁迫的重要候选基因。

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LI Li-shu, LUO Cong, AN Zhen-yu, LIU Zhao-liang, DONG Long, YU Hai-xia, HE Xin-hua. 2019. Molecular characterization, expression and function analysis of eukaryotic translation initiation factor (eIF1A) in Mangifera indica. Journal of Integrative Agriculture, 18(11): 2505-2513. 

Pre-harvest spraying of oxalic acid improves postharvest quality associated with increase in ascorbic acid and regulation of ethanol fermentation in kiwifruit cv. Bruno during storage

✺ Maratab Ali, LIU Meng-meng, WANG Zhen-e, LI Sheng-e, JIANG Tian-jia, ZHENG Xiao-lin  

The kiwifruit trees (Actinidia deliciosa cv. Bruno) were sprayed with 5 mmol L–1 oxalic acid (OA) or water (as control) at 130, 137 or 144 d after full-blossom, and then the fruit were harvested at commercial maturity and stored at room temperature (20±1)°C for 13 d. The effect of pre-harvest spraying of OA on postharvest quality of kiwifruit was evaluated during storage. The OA spraying slowed the increase in soluble solids content (SSC) and decrease in titratable acid (TA), as well as increased contents of ascorbic acid (AsA) and total-AsA accompanied with higher AsA/DHA ratio in kiwifruit during storage. Moreover, the OA spraying significantly reduced the contents of acetaldehyde and ethanol in kiwifruit, along with significant decrease in activities of enzymes involved in ethanol fermentation metabolism during the later period of storage, which was beneficial to control off-flavor associated with over accumulation of ethanol during postharvest. It was suggested that pre-harvest spraying of OA might maintain the postharvest quality of kiwifruit in relation to delay in fruit ripening, AsA maintenance and regulation of ethanol fermentation.

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Maratab Ali, LIU Meng-meng, WANG Zhen-e, LI Sheng-e, JIANG Tian-jia, ZHENG Xiao-lin. 2019. Pre-harvest spraying of oxalic acid improves postharvest quality associated with increase in ascorbic acid and regulation of ethanol fermentation in kiwifruit cv. Bruno during storage. Journal of Integrative Agriculture, 18(11): 2514-2520.

Modelling seedling development using thermal effectiveness and photosynthetically active radiation

利用辐热积模拟幼苗生长发育

✺ ZHOU Tian-mei, WU Zhen, WANG Ya-chen, SU Xiao-jun, QIN Chao-xuan, HUO He-qiang, JIANG Fang-ling

作物幼苗品质是其田间表现的先决条件，并且会影响其产量。温度和光照是育苗过程中影响秧苗品质的两个主要因素。苗床温度或光照不足，常会导致幼苗徒长或生产出瘦弱秧苗，造成严重的经济损失。若缺乏系统管理苗圃环境的知识，生产健康幼苗具有一定难度。本研究拟通过建立模拟模型来预测温度和光照协同作用对番茄和甘蓝幼苗生长的影响。首先，通过研究幼苗质量(根冠比、G值、壮苗指标)与辐热积的相关性，建立模型，并用独立的试验数据进行验证。结果表明，壮苗指数1 (HI1)和辐热积模型的曲线拟合良好，R²系数较高，说明该模型具有较高的可信度。番茄和甘蓝的HI1模拟模型分别为HI1=0.0009e^0.0308 TEP−0.0015和HI1=0.0003 e^0.0671 TEP−0.0003，可以用于预测在不同温度和光照条件下生长的番茄和甘蓝幼苗品质。↑ 向上滑动查看更多

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ZHOU Tian-mei, WU Zhen, WANG Ya-chen, SU Xiao-jun, QIN Chao-xuan, HUO He-qiang, JIANG Fang-ling. 2019. Modelling seedling development using thermal effectiveness and photosynthetically active radiation. Journal of Integrative Agriculture, 18(11): 2521-2533.

Plant Protection

Physicochemical properties and release characteristics of calcium alginate microspheres loaded with Trichoderma viride spores

✺ Slaven Jurić, Edyta Đermić, Snježana Topolovec-Pintarić, Marta Bedek, Marko Vinceković

Novel agroformulations for simultaneous delivery of chemical and biologically active agents to the plants were prepared by encapsulation of Trichoderma viride spores in calcium alginate microspheres. The impact of calcium ions concentration on the viability and sporulation of T. viride spores as well as on the microsphere important physicochemical properties were investigated. Intermolecular interactions in microspheres are complex including mainly hydrogen bonds and electrostatic interactions. T. viride germination inside matrix and germ tubes penetration out of microspheres revealed calcium alginate microspheres provide a supportive environment for T. viride growth. Differences in physicochemical properties and bioactive agents release behaviour from microspheres were ascribed to the changes in microsphere structure.  Fitting to Korsmeyer-Peppas empirical model revealed the underlying T. viride release mechanism as anomalous transport kinetics (a combination of two diffusion mechanisms and the Type II transport (polymer swelling and relaxation of the polymeric matrix)). The increasing amount of T. viride spores in the surrounding medium is closely related to the release from microspheres and germination.  The rate controlling mechanism of calcium release is Fickian diffusion. A decrease in the release rate with increasing calcium ion concentrations is in accordance with the calcium ions effect on the strength of the alginate network structure. T. viride germination inside microsphere diminished the amount of released calcium ions and slowed release kinetics in comparison with microspheres prepared without T. viride. The results indicated investigated agroformulations have a great potential to be used for plant protection and nutrition.↑ 向上滑动查看更多

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Slaven Jurić, Edyta Đermić, Snježana Topolovec-Pintarić, Marta Bedek, Marko Vinceković. 2019. Physicochemical properties and release characteristics of calcium alginate microspheres loaded with Trichoderma viride spores. Journal of Integrative Agriculture, 18(11): 2534-2548. 

Molecular, serological and biological characterization of a novel Apple stem pitting virus strain from a local pear variety grown in China

来源于中国一个地方梨品种的苹果茎痘病毒新株系的分子生物学及血清学特性研究

✺ LI Liu, ZHENG Meng-meng, MA Xiao-fang, LI Yuan-jun, LI Qing-yu, WANG Guo-ping, HONG Ni   

苹果茎痘病毒(Apple stem pitting virus，ASPV)是 β线性病毒科(Betaflexiviridae)凹陷病毒属(Foveavirus)的代表种。在自然条件下主要侵染苹果(Mulus spp.)和梨(Pyrus spp.)，在有的栽培梨上可引起叶脉黄化和果实石痘病。茌梨是中国山东莱阳及周边地区特有的地方梨品种，具较久的栽培历史，但目前尚未见有关当地保存的茌梨树的病毒鉴定及分子特性研究的报道。本研究采用逆转录PCR（RT-PCR）技术从莱阳地区栽培的近300年生的茌梨植株上检测到ASPV，通过设计引物进行RT-PCR扩增和PCR产物测序及序列拼接，首次获得了1个ASPV莱阳茌梨分离物（ASPV-LYC）的全基因组序列，ASPV-LYC基因组全长由9273个核苷酸组成（不包含 3'末端poly（A）尾），基因组结构与已报道的ASPV分离物相同。序列比对结果显示，ASPV-LYC与NCBI上登录的ASPV分离物全基因组序列相似性为73.1%-81.8%。遗传重组分析发现，ASPV-LYC的ORF1中存在一个重组区域，主要亲本和次要亲本分别为Aurora-1和KL9。对 ASPV-LYC的外壳蛋白（CP）进行原核表达，采用3个ASPV分离物（HB-HN6-8，HB-HN9-3和YN-MRS-17）的多克隆抗体进行western blot分析，结果显示ASPV-LYC与这3个ASPV分离物血清学相关，但在血清学反应活性上存在差异。进一步将ASPV-LYC嫁接传染至指示植物杂种榅桲（Pyronia veitchii）上，该分离物可引起杂种榅桲叶片反卷及茎干凹陷点等ASPV特有的症状。这些结果表明LYC为一个新的ASPV分离物，为深入了解ASPV的分子进化提供了重要信息。

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LI Liu, ZHENG Meng-meng, MA Xiao-fang, LI Yuan-jun, LI Qing-yu, WANG Guo-ping, HONG Ni. 2019. Molecular, serological and biological characterization of a novel Apple stem pitting virus strain from a local pear variety grown in China. Journal of Integrative Agriculture, 18(11): 2549-2560. 

Evaluation of the biocontrol potential of Aspergillus welwitschiae against the root-knot nematode Meloidogyne graminicola in rice (Oryza sativa L.) 

百岁兰曲霉生防真菌对水稻拟禾本科根结线虫的防治效果评价

✺ LIU Ying, DING Zhong, PENG De-liang, LIU Shi-ming, KONG Ling-an, PENG Huan, XIANG Chao, LI Zhong-cai, HUANG Wen-kun

拟禾本科根结线虫(Meloidogyne graminicola)是世界水稻产区危害最严重的病原线虫之一。由于化学杀线虫剂对环境危害较大且使用成本较高，我们必须筛选对环境有益且易于推广的生物防治制剂来替代化学农药。通过室内生测与温室盆栽实验，我们研究了百岁兰曲霉(Aspergillus welwitschiae)AW2017 对拟禾本科根结线虫的生防潜能。利用室内离体实验测定了AW2017对根结线虫卵孵化的抑制作用及其对幼虫的毒杀作用，利用温室盆栽实验研究了AW2017处理后水稻对线虫的诱集作用及线虫对水稻的侵入作用。结果表明，AW2017可以有效地抑制拟禾本科根结线虫卵的孵化且对幼虫有良好的毒杀作用。利用5×AW2017处理48 h后，幼虫死亡率达86.2%；利用5×AW2017处理7天后，卵孵化抑制率达47.3%。温室盆栽实验表明，5×AW2017可分别降低根结及根内侵入的线虫数量达 24.5%和40.5%，还可减少水稻根系对根结线虫的诱集作用、抑制侵入根内的线虫发育。因此，百岁兰曲霉代谢产物对拟禾本科根结线虫具有较好的防治效果，具有良好的开发应用前景。↑ 向上滑动查看更多

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LIU Ying, DING Zhong, PENG De-liang, LIU Shi-ming, KONG Ling-an, PENG Huan, XIANG Chao, LI Zhong-cai, HUANG Wen-kun. 2019. Evaluation of the biocontrol potential of Aspergillus welwitschiae against the root-knot nematode Meloidogyne graminicola in rice (Oryza sativa L.). Journal of Integrative Agriculture, 18(11): 2561-2570. 

Characterization of field-evolved resistance to cyantraniliprole in Bemisia tabaci MED from China

我国烟粉虱MED隐种对溴氰虫酰胺的田间抗性特征研究

✺ WANG Ran, WANG Jin-da, CHE Wu-nan, SUN Yan, LI Wen-xiang, LUO Chen

溴氰虫酰胺是新型的邻氨基苯甲酰胺类杀虫剂，当前利用其对世界性重要害虫烟粉虱的防治较为成功。基于前期已报道采自山西的田间抗性SX种群，本研究通过开展抗性衰退、抗性筛选、遗传分析、交互抗性以及增效作用等工作，进一步研究烟粉虱对溴氰虫酰胺的田间抗性特征。与敏感品系MED-S相比，SX种群的抗性水平为26.4倍。随后，利用SX种群建立了两个试验品系。其中，SX品系为不经筛选的抗性衰退品系，并且在实验过程中其抗性水平下降迅速且显著。另一个试验品系为SX-R品系，其经过连续多代的抗性筛选后，抗性水平已达到138.4倍。此外，结果显示SX-R品系对阿维菌素、吡虫啉、噻虫嗪、氟啶虫胺腈以及联苯菊酯不存在交互抗性，并且SX-R品系的抗性为常染色体不完全显性遗传。增效作用测定的结果显示，增效醚PBO对SX-R品系的抗性存在显著的增效作用。综上所述，经过对田间SX种群的连续多代筛选，成功建立了SX-R溴氰虫酰胺抗性品系，该品系抗性为不完全显性遗传且与田间常用的杀虫剂不存在交互抗性。此外，SX-R品系的抗性很可能与解毒代谢酶活力增强相关，但也存在靶标抗性的可能。

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WANG Ran, WANG Jin-da, CHE Wu-nan, SUN Yan, LI Wen-xiang, LUO Chen. 2019. Characterization of field-evolved resistance to cyantraniliprole in Bemisia tabaci MED from China. Journal of Integrative Agriculture, 18(11): 2571-2578..

Identification of diapause-associated proteins in migratory locust, Locusta migratoria L. (Orthoptera: Acridoidea) by label-free quantification analysis

利用无标记定量分析鉴定飞蝗滞育关联蛋白

✺ CUI Dong-nan, TU Xiong-bing, HAO Kun, Aftab Raza, CHEN Jun, Mark McNeill, ZHANG Ze-hua

飞蝗母本光周期反应是影响蝗卵滞育的重要因子，目前关于昆虫滞育已有较多报道，但是关于母本光周期反应影响后代滞育的调控机制尚未明确。为此，我们利用无标记定量分析了飞蝗滞育卵和非滞育卵蛋白组学数据。结果发现，滞育卵与非滞育卵中共有175个差异表达蛋白，与非滞育卵相比，滞育卵中显著上调的蛋白有24个、显著下调的蛋白有151个。GO和KEGG通路富集分析发现过氧化物酶代谢、昆虫激素生物合成、寿命调控等通路与飞蝗滞育调控关联。我们进一步利用qRT-PCR验证蛋白质组学分析的结果，发现昆虫储存蛋白4、保幼幼激素环氧化物水解酶1、细胞色素P450和热休克蛋白20.7等参与了滞育调控过程。本研究为进一步探索母本效应对后代滞育的调控提供了重要参考。

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CUI Dong-nan, TU Xiong-bing, HAO Kun, Aftab Raza, CHEN Jun, Mark McNeill, ZHANG Ze-hua. 2019. Identification of diapause-associated proteins in migratory locust, Locusta migratoria L. (Orthoptera: Acridoidea) by label-free quantification analysis. Journal of Integrative Agriculture, 18(11): 2579-2588. 

Animal Science · Veterinary Medicine

Two new lncRNAs regulate the key immune factor NOD1 and TRAF5 in chicken lymphocyte

✺ BI Yu-lin, YUAN Xiao-ya, CHEN Ying, CHANG Guo-bin, CHEN Guo-hong

Reticuloendotheliosis virus (REV) causes the atrophy of immune organs and immuno-suppression in chickens, but the underlying molecular mechanism of the immune response after infection by REV is not well understood.Presently, the RNA-seq was used to analyze the regulation of immune response to REV in chicken lymphocytes from peripheral blood. Overall, 134 differentially expressed long non-coding RNAs (lncRNAs) between cells with REV infection or no in vitro were screened. Based on the differentially expressed protein-coding genes, the NOD-like receptor pathway related to immune regulation was enriched. Two lncRNAs (L11530 and L09863) were predicted to respectively target the nucleotide-binding oligomerization domain 1 (NOD1) or tumor necrosis factor receptor-associated factor 5 (TRAF5) gene, which are involved in the NOD-like receptor pathway with cis-regulation way. The in vitro results revealed the significantly up-regulated (P<0.01) levels of lncRNA-L11530 and its target gene, NOD1, and the significantly down-regulated (P<0.05) levels of lncRNA-L09863 and its target gene, TRAF5 in lymphocytes after REV infection. These changes also occurred in vivo in blood lymphocytes of chickens infected with REV. Further, L09863 and L11530 were respectively interfered, the expression levels of their target genes NOD1 or TRAF5 had significantly down-regulated, accompanied by the change of IL-8 and IL-18 secretions in lymphocytes. The NOD-like receptor pathway appears to be important in the immune response to REV, LncRNA-11530 and lncRNA-09863 might involve in the immune regulation on REV infection by targeting NOD1 or TRAF5 in blood lymphocytes of chickens. Our findings reveal a new regulation of lncRNAs (L11530 and L09863) on immunity in chicken peripheral blood lymphocytes for REV infection by changing the expression of the target gene via the NOD-like receptor pathway.

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BI Yu-lin, YUAN Xiao-ya, CHEN Ying, CHANG Guo-bin, CHEN Guo-hong. 2019. Two new lncRNAs regulate the key immune factor NOD1 and TRAF5 in chicken lymphocyte. Journal of Integrative Agriculture, 18(11): 2589-2597.

Baculovirus-expressed FAdV-4 penton base protein protects chicken against hepatitis-hydropericardium syndrome

杆状病毒表达的禽腺病毒4型五邻体基质蛋白对鸡的免疫效果评价

✺ ZHANG Jun-qin, WEI Yan-ming, HUANG Kun, SUN Xiao-mei, ZOU Zhong, JIN Mei-lin

肝炎心包积液综合征(HHS)是由禽腺病毒血清4型(Fowl adenovirus serotype 4，FAdV-4)引起的一种传染病。近些年来，我国多个省份暴发了该病，给我国家禽养殖业造成了严重的经济损失。为防控该病，已有多项研究使用大肠杆菌表达系统或杆状病毒表达系统表达FAdV-4的多个免疫原性蛋白开发亚单位疫苗。然而，杆状病毒表达的五邻体基质蛋白(penton base)对鸡的免疫保护作用尚未评估。因此，本研究使用杆状病毒表达系统表达五邻体基质蛋白和纤突蛋白2(fiber-2)。随后使用镍柱纯化五邻体基质蛋白和纤突蛋白2，这两个蛋白分别免疫或共同免疫14日龄SPF鸡，28日龄加强免疫一次。42日龄时，FAdV-4强毒株攻毒免疫组和攻毒对照组。结果表明，单独免疫penton base或免疫penton base+fiber-2，攻毒保护效率100%；免疫fiber-2攻毒保护率90%，攻毒对照组保护率10%。所有接种重组蛋白的鸡均可产生抗体，剖检和病理切片检查显示免疫组组织无明显损伤，而攻毒对照组心脏和肝脏有禽腺病毒血清4型感染的典型病变。因此，杆状病毒表达的penton base蛋白能够对FAdV-4强毒株攻毒提供保护，是防治肝炎心包积液综合征候选亚单位疫苗，具有良好的应用前景。本研究首次利用杆状病毒表达系统高效表达五邻体基质蛋白，并评价杆状病毒表达五邻体基质蛋白对鸡的免疫效果。大团聚体形成与周转是影响土壤有机质动态变化及稳定的关键因素，同时团聚体粒径分布也与微生物群落组成和活性密切相关。

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ZHANG Jun-qin, WEI Yan-ming, HUANG Kun, SUN Xiao-mei, ZOU Zhong, JIN Mei-lin. 2019. Baculovirus-expressed FAdV-4 penton base protein protects chicken against hepatitis-hydropericardium syndrome. Journal of Integrative Agriculture, 18(11): 2598-2604.

Agro-Ecosystem & Environment

Soil macroaggregates and organic-matter content regulate microbial communities and enzymatic activity in a Chinese Mollisol

黑土大团聚体和有机质对土壤微生物群落和酶活性的影响

✺ CHEN Xu, HAN Xiao-zeng, YOU Meng-yang, YAN Jun, LU Xin-chun, William R. Horwath, ZOU Wen-xiu

本研究采取黑土区30余年长期定位试验处理下自然草地（GL）、农田（FL）和休耕裸地（BF）土壤进行60天室内培养，设置完整（IAD）和破坏大团聚体（RAD）处理，在添加外源葡萄糖、丙氨酸和无机氮条件下，研究土壤微生物活性和群落结构与团聚体粒级分布的关系。结果表明，经过60天培养，土壤中细菌和真菌磷脂脂肪酸（PLFA）含量最高值出现在第7天，而放线菌则在第15天表现最高。土壤酶活性与PLFA含量密切相关，且草地下土壤PLFA含量与酶活性均表现最高。在未添加外源养分条件下，草地RAD与IAD处理相比，土壤PLFA含量、葡萄糖苷酶（BG）和乙酰葡萄胺糖苷酶（NAG）活性平均提高了15.7%、17.4%和7.6%。FL和BF土壤受外加碳源显著影响，土壤PLFA含量较未添加处理分别增加了18.8%和33.4%。团聚体分布对土壤有机碳稳定性和微生物群落结构及活性具有重要影响，大团聚体对有机碳的物理保护是土壤固碳的重要机制之一。

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CHEN Xu, HAN Xiao-zeng, YOU Meng-yang, YAN Jun, LU Xin-chun, William R. Horwath, ZOU Wen-xiu. 2019. Soil macroaggregates and organic-matter content regulate microbial communities and enzymatic activity in a Chinese Mollisol. Journal of Integrative Agriculture, 18(11): 2605-2618.

Assessment of the contribution percentage of inherent soil productivity of cultivated land in China

我国耕地基础地力贡献率的评价

✺ WANG Shi-chao, WANG Jin-zhou, ZHAO Ya-wen, REN Yi, XU Ming-gang, ZHANG Shu-xiang, LU Chang-ai

【目的】耕地基础地力贡献率是反应耕地土壤肥力状况的重要指标，了解我国耕地基础地力贡献率的现状与演变规律，对于我国耕地质量管理建设及提出耕地培育改良措施提供重要的科学依据。【方法】基于我国1984-1990年(简称“1980s”)、1996-2000年(简称“1990s”)、2004-2013年(简称“2000s”)三个时段建成国家耕地质量监测点和27个长期定位试验（1979-2013年）的监测资料，研究我国水稻、小麦和玉米三大粮食作物的耕地基础地力贡献率、区域耕地地力的演变规律。【结果】目前我国三大粮食作物的耕地基础地力贡献率53.8%。1990s年代（57.5%）我国三大作物的基础地力贡献率显著高于1980s（50.3%）和1990s年代（52.0%）；连续多年不施肥后，耕地地力贡献率呈逐渐下降的趋势，并在一定时期后达到较为稳定的状态，其中西北和东北一年一熟制旱地地力贡献率呈下降趋势，平均每年降低4.5%和1.0%，10年后逐渐趋于稳定；而北方旱一年两熟制旱地地力贡献率呈逐年下降趋势，下降速率为1.1%；水旱轮作制地力贡献率略有下降（0.07%），但水田耕地地力贡献率显著下降（3.1%），至2年后趋于稳定。综上，复种指数高的轮作制度对旱地地力的消耗更为严重，而水田基础地力贡献率维持较高水平。【结论】为了发挥我国作物新品种的增产潜力，需采取措施提高我国的耕地基础地力，为作物高产稳产奠定坚实基础。↑ 向上滑动查看更多

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WANG Shi-chao, WANG Jin-zhou, ZHAO Ya-wen, REN Yi, XU Ming-gang, ZHANG Shu-xiang, LU Chang-ai. 2019. Assessment of the contribution percentage of inherent soil productivity of cultivated land in China. Journal of Integrative Agriculture, 18(11): 2619-2627.

Winter wheat identification by integrating spectral and temporal information derived from multi-resolution remote sensing data

综合多分辨率遥感数据光谱和时间信息的冬小麦识别研究

✺ ZHANG Xi-wang, LIU Jian-feng, Zhenyue Qin, QIN Fen

及时的作物面积和分布信息是驱动农业相关应用的基础数据。对于基于遥感的作物类型识别，仅利用单一类型影像的方法有明显的局限性。当前，综合精细分辨率和粗分辨率遥感影像的研究，如像元分解、回归模型，等，通常产生粗分辨率的丰度图，没有像元内的细节信息。很少有研究在识别中考虑这两类影像像元之间的空间关系。本文提出了一种新的冬小麦识别方法，通过综合多分辨率遥感数据的光谱和时间信息，确定粗分辨率像元内子像元的空间分布。首先，基于贝叶斯公式利用重采样得到的25米分辨率TM数据确定冬小麦的隶属度。然后，基于来源于MODIS时间序列数据的冬小麦独特的时间变化特征，利用多元回归模型估算冬小麦丰度。最后，利用本文提出的AM（abundance-membership）模型识别冬小麦，该模型基于两种分辨率遥感数据的像元间的空间关系。具体来说，通过比较每个粗分辨率丰度像元空间上所对应的10×10个隶属度像元，从而识别冬小麦。换句话说，该方法利用了在局部范围内的隶属度的像对大小，而不是整个研究区内的绝对大小。该方法在伊洛河流域主要农业区进行试验，结果显示面积精度（Aa）为93.01%，采样精度（As）为91.40%。混淆矩阵结果显示，总体精度（OA）为91.40%，Kappa系数为0.755。相对于传统仅依赖光谱特征的最大似然法分类和随机森林分类方法，精度明显得到提高。研究结果说明，通过综合光谱和时间信息可以提高识别精度。由于该方法识别冬小麦是在对应于每一个MODIS像元大小的特定空间内进行识别，因此环境条件差异的影响也被明显降低。这个优势将使该方法可以有效地应用在其他地区。↑ 向上滑动查看更多

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ZHANG Xi-wang, LIU Jian-feng, Zhenyue Qin, QIN Fen. 2019. Winter wheat identification by integrating spectral and temporal information derived from multi-resolution remote sensing data. Journal of Integrative Agriculture, 18(11): 2628-2643.

Food Science

Changes of moisture distribution and migration in fresh ear corn during storage

贮藏期间新鲜穗玉米水分分布和迁移的变化

✺ WANG Hao, LIU Jing-sheng, MIN Wei-hong, ZHENG Ming-zhu, LI Hao

为了保持新鲜穗玉米的质量，必须了解其在储存过程中的水分变化。本项研究利用低场核磁共振和磁共振成像研究了新鲜穗玉米在储存过程中水分分布和迁移的变化。结果表明，在储存的第一个小时内，新鲜穗玉米中的水分损失大于水分迁移。此后随着储存时间的增加，水分损失比水分迁移弱。随着贮藏时间的延长，切片新鲜玉米芯的低场核磁共振信号强度呈下降趋势，外围区域的信号强度降低率高于中央部位。当贮藏的新鲜穗玉米中总水分含量降低时，结合水的相对比例增加而游离水的比例则下降。总而言之，低场核磁共振和磁共振成像是用于实时监测新鲜穗玉米在存储过程中水分分布，迁移和损失以评估其质量的有效且无损的工具。这些结果可用于将来设计新鲜穗玉米的保存和加工条件。↑ 向上滑动查看更多

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WANG Hao, LIU Jing-sheng, MIN Wei-hong, ZHENG Ming-zhu, LI Hao. 2019. Changes of moisture distribution and migration in fresh ear corn during storage. Journal of Integrative Agriculture, 18(11): 2644-2651.

Relationship between physicochemical characteristics of Korean wheat flour and quality attributes of steamed bread

✺ Ji-Eun Kim, Byung-Kee Baik, Chul Soo Park, Jae-Han Son, Chang-Hyun Choi, Youngjun Mo, Tae-Il Park, Chon-Sik Kang, Seong-Woo Cho

The purpose of this study is to identify major factors affecting the manufacture and quality of steamed bread, consumed in Southeast Asia including China, Japan, and Korea. Hence, flours of 11 Korean wheat cultivars were used to evaluate quality attributes of two different styles of steamed bread, Korean style steamed bread (KSSB) and northern-style Chinese steamed bread (NSCSB). KSSB prepared more ingredients and higher optimum water absorption of dough than NSCSB because Korean consumers prefer white and glossy surface and soft crumb. KSSB showed lower height, larger diameter and volume of steamed bread, higher stress relaxation, and softer texture of crumb than NSCSB. The correlation between flour characteristics and quality of steamed bread was different in KSSB and NSCSB.  About 90% of variability in the height and volume of KSSB could be predicted from protein content, mixing tolerance of Mixograph, average particle size of flour, final viscosity and solvent retention capacity. Protein content and quality parameters also could explain the variation of steamed bread height in NSCSB.  Korean wheat carrying Glu-A3c allele produced higher volume of steamed bread (704.7 mL) than Glu-A3d allele (645.8 mL) in KSSB, although there was no significant difference in volume of NSCSB by glutenin compositions.  Glu-D1d and Glu-A3c alleles had softer texture of crumb than Glu-D1f and Glu-A3d alleles in KSSB, Glu-B3i allele also showed lower hardness of crumb than their counterpart allele in NSCSB. Hard wheat showed higher height and volume of steamed bread, and lower stress relaxation and hardness of crumb than soft wheat in KSSB.

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Ji-Eun Kim, Byung-Kee Baik, Chul Soo Park, Jae-Han Son, Chang-Hyun Choi, Youngjun Mo, Tae-Il Park, Chon-Sik Kang, Seong-Woo Cho. 2019. Relationship between physicochemical characteristics of Korean wheat flour and quality attributes of steamed bread. Journal of Integrative Agriculture, 18(11): 2652-2663

Short Communication

First detection and complete genome of chlorotic mottle virus naturally infecting soybean in China by deep sequencing

大豆褪绿斑驳病毒在我国的首次发现及其基因组全序列

✺ HU Qian-qian, LIU Xue-jian, HAN Xue-dong, LIU Yong, JIANG Jun-xi, XIE Yan

利用高通量测序技术在我国江西省的大豆上首次发现大豆褪绿斑驳病毒(Soybean chlorotic mottle virus, SbCMV)，并通过PCR验证。SbCMV是花椰菜花叶病毒科大豆褪绿斑驳病毒属病毒的典型代表种，目前仅在日本报道。序列测定发现，我国NC113分离物的全基因组为8210个核苷酸(基因登录号：MH718847)，与日本报道的SbCMV-JA全序列同源性最高为91.7%。它含有典型的SbCMV基因组结构，为环状双链DNA分子，编码9个可能的读码框(Ia, Ib, II-VIII)，并在读码框VI和VII之间，核苷酸5976-6512处包含一个长为537个核苷酸的非编码区。对NC113的全基因组核苷酸序列与其编码的各蛋白的氨基酸序列进行比较分析发现，除与日本的SbCMV-JA同源性最高外，NC113与同属的蓝莓红环斑病毒(Blueberry red ringspot virus, BRRSV)、花生褪绿条纹病毒(Peanut chlorotic streak virus, PCSV)及夜香树黄曲叶病毒(Cestrum yellow leaf curling virus, CmYLCV)的全基因组序列同源性分别为53.8%、48.7%、47.6%，而与花椰菜花叶病毒科的其他5个属的代表种病毒的同源性都在40.3%以下，说明NC113是SbCMV的一个分离物。系统关系树分析表明SbCMV-NC113与同属的SbCMV、BRRSV、PCSV、CmYLCV亲缘关系最近聚在一起，而与花椰菜花叶病毒科的其他5个属的花椰菜花叶病毒(Cauliflower mosaic virus, CaMV)、木薯脉花叶病毒(Cassava vein mosaic virus, CsVMV)、鸭跖草黄色斑驳病毒(Commelina yellow mottle virus, ComYMV)、碧冬茄脉明病毒(Petunia vein clearing virus, PVCV)、水稻东格鲁杆状病毒(Rice tungro bacilliform virus, RTBV)亲缘关系较远各自形成独立的分支。对浙江省和江西省472个田间豆科植物样本进行PCR检测，仅在南昌地区的5株大豆上检测到特异条带。这是SbCMV在我国的首次报道。虽然目前SbCMV的发生率较低，但由于SbCMV易于机械传播，而其田间传毒介体不明，因此有必要在全国范围内对SbCMV的发生分布与危害作进一步研究。↑ 向上滑动查看更多

Cite this article:   

HU Qian-qian, LIU Xue-jian, HAN Xue-dong, LIU Yong, JIANG Jun-xi, XIE Yan. 2019. First detection and complete genome of chlorotic mottle virus naturally infecting soybean in China by deep sequencing. Journal of Integrative Agriculture, 18(11): 2664-2667.

Complete genome sequence of Bacillus amyloliquefaciens YP6, a plant growth rhizobacterium efficiently degrading a wide range of organophosphorus pesticides

一株高效降解多种有机磷农药的根际促生菌解淀粉芽孢杆菌YP6的全基因组测序

✺ MENG Di, ZHAI Li-xin, TIAN Qiao-peng, GUAN Zheng-bing, CAI Yu-jie, LIAO Xiang-ru

解淀粉芽孢杆菌YP6是一株能够高效降解多种有机磷农药的根际促生菌。本研究通过高通量测序报道了该菌株的全基因组，研究发现该菌株整个基因组大小为4009619bp，GC含量为45.9%，编码蛋白的基因总数为4210个。通过对其全基因组进行分析，我们发现该菌基因组中含有一些与溶磷、降解有机磷农药以及合成生长素和铁载体的相关基因。与解淀粉芽孢杆菌属中其他菌株的全基因组对比结果显示：该菌株基因组更长，编码蛋白的基因更多；但是参与细胞壁/细胞膜/包膜合成（M）、翻译，核糖体结构及合成（J）、转录（K）和信号转导机制（T）的蛋白数量最少；这些差异可能与解淀粉芽孢杆菌属对不同环境的适应能力相关。通过全基因组测序与比较分析，我们初步发掘了YP6在环境生物修复中的应用潜力，为生物技术和基因工程提供了有机磷农药降解的基因资源，有助于我们更好的了解微生物与生存环境之间的关系。

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Cite this article:   

MENG Di, ZHAI Li-xin, TIAN Qiao-peng, GUAN Zheng-bing, CAI Yu-jie, LIAO Xiang-ru. 2019. Complete genome sequence of Bacillus amyloliquefaciens YP6, a plant growth rhizobacterium efficiently degrading a wide range of organophosphorus pesticides. Journal of Integrative Agriculture, 18(11): 2668-2672. 

Journal of Integrative Agriculture (《农业科学学报》，JIA) 杂志是中国农业科学院与中国农学会联合主办、农业信息研究所承办的英文学术期刊。创刊于2002年，主编为中国农业科学院副院长万建民院士。2018年SCI影响因子1.337，位于科睿唯安JCR农业综合学科Q2区（18/56）。2019年采用开放获取形式，专注刊登作物学、园艺、植物保护、动物科学、动物医学、资源环境、食品科学、农业经济与管理等国际农业科学前沿热点领域的综述、研究论文、简报以及评述等。

 

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