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1. JIA-2021-0354 利用线粒体D_Loop序列研究全世界山羊母系遗传多样性及系统发育分析
GUO Yi, GONG Ying, HE Yong-meng, YANG Bai-gao, ZHANG Wei-yi, CHEN Bo-er, HUANG Yong-fu, ZHAO Yong-ju, ZHANG Dan-ping, MA Yue-hui, CHU Ming-xing, E Guang-xin
Journal of Integrative Agriculture    2022, 21 (6): 1830-1837.   DOI: 10.1016/S2095-3119(21)63882-0
摘要244)      PDF    收藏

本研究利用已公布的4165个来自于全世界196个品种的山羊线粒体D_Loop序列,进行核苷酸多样性、单倍型构建、单倍型多样性、群体系统发育学研究、中性检验及群体遗传距离等一系列遗传参数进行评估。在全部个体的401 bp片段长度的D_Loop区域内共鉴定得到301个多态位点,总体核苷酸多样性为0.03471;构建获得并构建2409个D_Loop单倍型,单倍型平均多样性为0.9983。系统发育分析表明,98.92%的单倍型被聚类为已知的6个山羊线粒体D_loop单倍型簇,其中单倍型A所占比例最大(86%),D_Loop单倍型B簇在中国山羊中出现频率最高。中国西南地区山羊群体中发现了两个未知的D_Loop单倍型簇(Unknown I和Unknown II)。分子方差分析(AMOVA)和群体间成对差异(PiXY)研究结果表明,不同品种家养山羊间群体变异较小,群体遗传分化与地理分布不完全一致,表明群体间广泛存在遗传物质交流。中性检验(Tajima‘D和Fu’Fs检验)和错配分布研究结果表明,单倍型簇B、C和G存在群体扩张历史。较其他野羊,Capra aegagrus与家养山羊系统发育关系最为密切,并可能贡献于山羊A、B、C和F单倍群簇的驯化起源。本研究表明线粒体D_Loop单倍型B簇可能起源于中国或在山羊驯化早期已迁至中国,在中国西南地区山羊群体中两个未知的D_Loop单倍型簇的发现表明中国西南地区可能具有独特的山羊母系背景或驯化历史;Capra aegagrus是最可能的家养山羊野生祖先,并可能贡献于山羊A、B、C和F单倍群簇的驯化起源。本研究利用大样本量全世界山羊线粒体D_Loop序列数据集分析有助于更好的理解世界范围内山羊母系驯化起源及基因流动的历史变化,为进一步明确世界山羊群体迁徙的演变历史及种群系统发育定位提供了宝贵的理论依据。


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2. JIA-2021-1450 一种基于腋芽离体再生技术的柑橘黄龙病菌快速增殖系统的构建
LEI Tian-gang, HE Yong-rui, ZOU Xiu-ping, WANG Xue-feng, FU Shi-min, PENG Ai-hong, XU Lan-zhen, YAO Li-xiao, CHEN Shan-chun, ZHOU Chang-yong
Journal of Integrative Agriculture    2022, 21 (6): 1683-1693.   DOI: 10.1016/S2095-3119(21)63856-X
摘要198)      PDF    收藏

本研究以感染黄龙病菌亚洲种(‘Candidatus Liberibacter asiaticus’, CLas)的长叶橙(Citrus sinensis Osbeck)为材料,建立CLas侵染长叶橙的茎段培养方法,利用PCR和荧光定量PCR检测分析腋芽离体再生嫩梢中CLas的增殖情况。在此基础上,取CLas侵染长叶橙枝条进行表面消毒处理,切取腋芽嫁接于柑橘试管砧木,进行试管培养,利用PCR和荧光定量PCR检测腋芽再生嫩梢中CLas的增殖规律;采用直接组织印迹免疫法(Direct tissue blot immune assay,DTBIA)对试管苗中CLas的分布进行检测。培养基中添加适合浓度的激素可促进感病柑橘茎段离体培养的萌芽,萌芽率最高的培养基为MS+6-BA 1.0 mg·L-1+GA 0.2 mg·L-1+IAA 0.2 mg·L-1。茎段萌芽30 d时,嫩梢病原PCR检测的阳性率达75%,其CLas浓度平均为温室条件下培养茎段原叶片中脉的28.2倍,最高为484.2倍。试管嫁接苗萌芽10、15、20、25、30和40 d时,嫩梢CLas PCR检测的阳性率分别为10%、15%、15%、20%、55%和70%,CLas浓度分别为7.5×104 、2.2×106 、1.4×107、2.2×107 、1.2×108和1.4×108 cells μg-1 DNA;萌芽30和40 d的试管苗中CLas浓度超过108的比例分别达30%和40%。DTBIA检测结果显示,试管苗中黄龙病菌的分布比温室保存的感病柑橘中均匀。CLas侵染柑橘试管苗的主要症状为嫩梢枯死、停止生长、叶黄和落叶。感病试管苗的死亡率在萌芽40 d后会急剧升高,60 d时死亡率达82.0%。CLas在通过腋芽嫁接的柑橘试管苗中能快速增殖,该方法的建立为深入开展柑橘黄龙病病原生物学、病原-寄主互作及抗菌药物快速筛选等研究奠定一定基础。本研究建立了一种快速增殖和高浓度富集柑橘黄龙病菌的培养方法


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3. 快速,低成本的深度学习系统可基于云服务对草莓病害进行分类
YANG Guo-feng, YANG Yong, HE Zi-kang, ZHANG Xin-yu, HE Yong
Journal of Integrative Agriculture    2022, 21 (2): 460-473.   DOI: 10.1016/S2095-3119(21)63604-3
摘要179)      PDF    收藏

准确、及时地对草莓种植过程中的病害进行分类,可以帮助种植者对其进行及时的处理,从而减少损失。但真实种植环境下的草莓病害分类面临着严峻的挑战,包括复杂的种植环境,多种差异较小的病害类别等。尽管最近基于深度学习的移动视觉技术在克服上述问题方面取得了一些成功,但对多地域、多空间、多时间的草莓病害分类需求而言,一个关键问题是如何构建一种无损、快速、便捷的方法提高草莓病害识别的效率。我们开发并评估一种快速,低成本的系统,用于对草莓种植中的病害进行分类。这涉及设计一个易于使用的基于云服务的草莓病害识别系统,并结合我们提出的新颖的自监督多网络协作的分类模型,它由定位网络、反馈网络和分类网络组成,以识别草莓常见病害的类别。该模型借助新颖的自我监督机制,可以有效地识别草莓病害图像中的病害区域,而不需要边界框等标注。使用准确率,精确率,召回率和值来评估分类效果,测试集的结果分别为92.48%,90.68%,86.32%和88.45%。与流行的卷积神经网络和其他五种方法相比,该网络实现了更好的病害分类效果。目前,系统的客户端(小程序)已上线微信平台。小程序在实际测试中分类效果良好,验证了系统的可行性和有效性,能够为草莓病害识别的智能化研究与应用提供参考。

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4. Fluorescent competitive assay for melamine using dummy molecularly imprinted polymers as antibody mimics
DU Xin-wei, ZHANG Yan-xin, SHE Yong-xin, LIU Guang-yang, ZHAO Feng-nian, WANG Jing, WANG Shan-shan, JIN Fen, SHAO Hua, JIN Mao-jun, ZHENG Lu-fei
Journal of Integrative Agriculture    2016, 15 (05): 1166-1177.   DOI: 10.1016/S2095-3119(16)61357-6
摘要2482)      PDF    收藏
    A fluorescent competitive assay for melamine was first developed utilizing dummy molecularly imprinted polymers (DMIPs) as artificial antibodies. This method is based on the competition between fluorescent substances and the unlabeled analyte for binding sites in synthesized DMIPs and the decreased binding of fluorescent substances to DMIPs due to increased concentrations of melamine in the solutions. DMIPs for melamine were synthesized under a hot water bath in the presence of the initiator azobisisobutyronitrile (AIBN) using 2,4-diamino-6-methyl-1,3,5-triazine (DAMT) as a dummy template, methacrylic acid (MAA) as a functional monomer, and ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent. The adsorption capacity and selectivity of DMIPs for melamine were evaluated by the isothermal adsorption curve and Scatchard analysis. The evaluation results showed that the synthesized DMIPs had specific recognition sites for melamine and the maximum adsorption amount was 1 066.33 μg g–1. Later, 5-(4,6-dichlorotriazinyl) amino fluorescein (DTAF) with a triazine ring, which slightly resembles melamine, was selected as the fluorescent substance. The fluorescent competitive assay using DMIPs as the antibody mimics was finally established by selecting and optimizing the reaction solvents, DMIPs amount, DTAF concentration, and incubation time. The optimal detection system showed a linear response within range of 0.05–40 mg L–1 and the limit of detection (LOD) was 1.23 μg L–1. It was successfully applied to the detection of melamine in spiked milk samples with satisfactory recoveries (71.9 to 86.3%). According to the comparative analysis, the result of optimized fluorescent competitive assay revealed excellent agreement with the HPLC-MS/MS result for melamine.
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5. Low Root Zone Temperature Exacerbates the Ion Imbalance and Photosynthesis Inhibition and Induces Antioxidant Responses in Tomato Plants Under Salinity
HE Yong, YANG Jing, ZHU Biao , ZHU Zhu-jun
Journal of Integrative Agriculture    2014, 13 (1): 89-99.   DOI: 10.1016/S2095-3119(13)60586-9
摘要1979)      PDF    收藏
The combined effects of salinity with low root zone temperature (RZT) on plant growth and photosynthesis were studied in tomato (Solanum lycopersicum) plants. The plants were exposed to two different root zone temperatures (28/20°C, 12/8°C, day/night temperature) in combination with two NaCl levels (0 and 100 mmol L-1). After 2 wk of treatment, K+ and Na+ concentration, leaf photosynthetic gas exchange, chlorophyll fluorescence and leaf antioxidant enzyme activities were measured. Salinity significantly decreased plant biomass, net photosynthesis rate, actual quantum yield of photosynthesis and concentration of K+, but remarkably increased the concentration of Na+. These effects were more pronounced when the salinity treatments were combined with the treatment of low RZT conditions. Either salinity or low RZT individually did not affect maximal efficiency of PSII photochemistry (Fv/Fm), while a combination of these two stresses decreased Fv/Fm considerably, indicating that the photo-damage occurred under such conditions. Non-photochemical quenching was increased by salt stress in accompany with the enhancement of the de-epoxidation state of the xanthophyll cycle, in contrast, this was not the case with low RZT applied individually. Salinity stress individually increased the activities of SOD, APX, GPOD and GR, and decreased the activities of DHAR. Due to the interactive effects of salinity with low RZT, these five enzyme activities increased sharply in the combined stressed plants. These results indicate that low RZT exacerbates the ion imbalance, PSII damage and photosynthesis inhibition in tomato plants under salinity. In response to the oxidative stress under salinity in combination with low RZT, the activities of antioxidant enzymes SOD, APX, GPOD, DHAR and GR were clearly enhanced in tomato plants.
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