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1. 挥发性代谢组和转录组揭示大花粉晕香水月季花香释放节律及分子机制
ZHOU Li-jun, HUANG Run-huan, LIU Ting-han, LIU Wei-chao, CHEN Yun-yi, LU Pei-feng, LUO Le, PAN Hui-tang, YU Chao, ZHANG Qi-xiang
Journal of Integrative Agriculture    2023, 22 (7): 2111-2125.   DOI: 10.1016/j.jia.2023.06.015
摘要263)      PDF    收藏

月季是世界上最重要的观赏植物之一,极高食用和药用价值,还全世界范围广泛种植用来提取精油。本研究采用固相微萃取(SPME)和气相色谱-质谱(GC-MS)联用法对在云南西北地区发现的具有甜香的新物种——大花粉晕香水月季(Rosa yangii不同开花阶段的花香成分进行提取和分析。共检测到113种挥发性有机化合物,从中筛选出69芳香挥发物我们发现大花粉晕香水月季的花朵在初开期产生和释放挥发性有机物明显高于其他开放阶段阶段合成并保留了大量的花香组分说明在工业生产更适合在初开期采收花朵。气味活性值(OAV)分析表明,大花粉晕香水月季的主要芳香成分包括丁香酚、甲基丁香醇、苯乙醛和苯乙醇、庚醛、癸醛、(E-2-己烯-1-基乙酸酯、石竹烯等。代谢组和时序基因共表达网络(TO-GCN联合分析显示,苯类/苯丙类挥发性有机化合物合成途径上的基因和有机挥发物其浓郁甜香起主要调控作用。MYBbHLH可能是调控丁香酚合成酶(EGS)异丁香酚合成酶(IGS)合成进而影响花香重要转录因子。综上所述,本研究可为观赏植物芳香分子育种提供科学依据,并促进植物精油新原料在食品储藏、芳香疗法、化妆品和香料行业的开发利用。

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2. PnSCR82, a small cysteine-rich secretory protein of Phytophthora nicotianae, can enhance defense responses in plants
WANG Ya-di, LI Fei, ZHANG Xin, LIU Ting-li, LIANG Wen-xing, LI De-long
Journal of Integrative Agriculture    2022, 21 (3): 751-761.   DOI: 10.1016/S2095-3119(21)63681-X
摘要187)      PDF    收藏

植物病原菌侵染植物过程中分泌大量的外泌蛋白调控植物免疫反应,这些外泌蛋白中包含一类富含半胱氨酸(small cysteine-rich, SCR)的小分泌蛋白,它能帮助病原菌在寄主植物中定殖。烟草疫霉中同样存在富含半胱氨酸外泌蛋白,但其功能尚不清楚。本研究通过生物信息学分析,发现烟草疫霉PnSCR82同源于恶疫霉的植物毒性蛋白PcF (Phytophthora cactorum Fragaria),其具有外泌信号肽,同时富含半胱氨酸。为研究烟草疫霉PnSCR82功能,将PnSCR82构建于马铃薯X病毒(PVX)表达载体(pGR106),利用农杆菌介导的转化方法注射本氏烟草叶片和番叶片茄,发现在烟草叶片瞬时表达PnSCR82,能够诱导烟草叶片和番茄叶片出现过敏性坏死,并且能增加胼胝质和活性氧在叶片的积累,接种辣椒疫霉后发现其增强对辣椒疫霉的抗性。通过构建其亚细胞定位载体PnSCR82-GFP,利用农杆菌介导的转化注射烟草叶片发现其定位于烟草细胞膜。同时RT-PCR分析发现在烟草叶片中瞬时表达PnSCR82能够激活烟草水杨酸,茉莉酸和乙烯途径的部分相关基因上调表达。烟草疫霉PnSCR82作为激发子一员,能够诱导烟草细胞过敏性坏死同时也能激活不同免疫信号通路相关基因上调表达,能够增强植物对辣椒疫霉的抗性,具有潜在的提高植物免疫应用价值。本文首次验证了烟草疫霉富含半胱氨酸蛋白PnSCR82功能,作为激发子,PnSCR82能够诱导植物产生过敏性坏死反应,能够激活部分免疫相关基因的上调表达,具有潜在的应用价值。


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3. 小麦DnaJ家族的全基因组鉴定及其对小麦黄花叶病毒侵染的调控分析
LIU Ting-ting, XU Miao-ze, GAO Shi-qi, ZHANG Yang, HU Yang, JIN Peng, CAI Lin-na, CHENG Ye, CHEN Jian-ping, YANG Jian, ZHONG Kai-li
Journal of Integrative Agriculture    2022, 21 (1): 153-169.   DOI: 10.1016/S2095-3119(21)63619-5
摘要258)      PDF    收藏

DnaJ蛋白最初是在大肠杆菌中鉴定的一种大小约为41kDa的热休克蛋白,其蛋白家族是分子伴侣中最多样化的家族,在蛋白质折叠和各种生理活动的调节中扮演了重要角色,且在植物发育和胁迫应答中发挥重要作用。DnaJ家族蛋白已在许多物种中广泛研究,例如人类,果蝇,蘑菇,西红柿和拟南芥等,但在小麦中的作用以及其与植物病毒之间的相互作用机制却鲜少有研究。在这篇文章中,我们鉴定了236TaDnaJs,并对其保守结构域,基因结构蛋白质基序蛋白质结构染色体定位和共线性以及顺式作用元件进行了全基因组分析。根据分析结果,我们将这些TaDnaJs按其结构域分DJADJBDJC三组,并从分组中随机选择了6个基因进行组织特异性分析和激素胁迫下的基因表达谱分析,结果表明TaDnaJ基因在不同组中存在组织差异表达,DJA组的基因表达集中顶部叶片,对ABAGA更为敏感;DJB组的基因表达水平在根和种子中最高,对ABA更为敏感;DJC组中的基因表达在小麦叶片中最高,其次是根和种子,对SAGA最敏感。另外我们随机选择了17个基因分析植物病毒侵染后基因表达水平的变化,结果显示,在测试的17TaDnaJ基因中,有16个基因在小麦黄花叶病毒(WYMV染后呈现上调表达,这表明TaDnaJ家族可能参与了植物防御反应。随后我们通过酵母两杂交实验验证了WYMV NIaNIb7KD蛋白与WYMV染后表达水平变化最显著的TraesCS7A02G506000相互作用。在这篇文章中,我们探究了DnaJ蛋白介导的胁迫耐受性和敏感性的分子机制,DnaJ基因可能参与了植物对非生物和生物胁迫的抗性。本研究提高了对TaDnaJ基因表达谱认识,并且为TaDnaJ家族与植物防御机制之间的研究提供了一定的研究基础。

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4. Transcriptomic insights into growth promotion effect of Trichoderma afroharzianum TM2-4 microbial agent on tomato plants
ZHAO Juan, LIU Ting, LIU Wei-cheng, ZHANG Dian-peng, DONG Dan, WU Hui-ling, ZHANG Tao-tao, LIU De-wen
Journal of Integrative Agriculture    2021, 20 (5): 1266-1276.   DOI: 10.1016/S2095-3119(20)63415-3
摘要157)      PDF    收藏

植物促生真菌具有产生生物活性物质、促进植物生长以及增强植物免疫抗性的能力,其作为有应用价值的有益微生物在作物栽培中受到越来越多关注。本研究从健康番茄植株根际土壤中分离筛选到一株木霉菌株TM2-4,将其鉴定为非洲哈茨木霉(Trichoderma afroharzianum)。菌株TM2-4发酵滤液中含有多种生物活性物质且对番茄种子发芽具有明显促进作用,发酵滤液100倍稀释液处理,番茄胚轴、胚根长度,种子活力指数分别增加28.7%、19.4% 和 62.1%。为了评价菌株TM2-4的促生作用及其相关机制,通过盆栽试验和转录组测序分析了非洲哈茨木霉菌制剂TM2-4处理对番茄植株生物学指标和基因表达型的影响。结果表明,非洲哈茨木霉TM2-4能够通过在植株根际土壤和根系有效定殖,显著提高番茄株高、干重、单株叶片数及根系活力等生物学指标。转录组分析发现,木霉菌制剂处理番茄根系较对照共获得984个差异表达基因,主要集中在激素平衡、抗氧化活性以及苯丙烷类生物合成和谷胱甘肽代谢等生物学过程。相关研究结果为阐明菌株TM2-4对番茄的促生作用机制提供有效信息,并为非洲哈茨木霉微生物菌剂在蔬菜作物生产中的进一步开发应用奠定理论基础。


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5. Using proximal sensor data for soil salinity management and mapping
GUO Yan, ZHOU Yin, ZHOU Lian-qing, LIU Ting, WANG Lai-gang, CHENG Yong-zheng, HE Jia, ZHENG Guo-qing
Journal of Integrative Agriculture    2019, 18 (2): 340-349.   DOI: 10.1016/S1671-2927(00)12104
摘要260)      PDF(pc) (5151KB)(548)    收藏
Over the past five decades, increased pressure caused by the rapidly growing population has resulted in a reclamation of agricultural and urban buffer zones along China’s coastline.  However, information about the spatio–temporal variation of soil salinity in these reclaimed regions is limited.  As such, obtaining this information is crucial for mapping the variation in saline areas and to identify suitable salinity management strategies.  In this study, we employed EM38 data to conduct digital soil mapping of spatio–temporal variation and map these variations of different site-specific zones.  The results indicated that the distribution of soil salinity was heterogeneous in the middle of, and that the leaching of salts was significant at the edges of, the study field.  Afterwards, fuzzy-k means algorithm was used to divide the site-specific management zones within the time series apparent soil electrical conductivity (ECa) data and the spatial correlations of variation.  We concluded that two management zones are optimal to guide precision management.  Zone A had an average salinity level of about 165 mS m–1, in which salt-tolerant crops, such as cotton and barley can grow normally, while crops such as soybean and cowpeas may be planted using leaching and increasing the mulching film methods to reduce the accumulation of salt in surface soil.  In Zone B, there was a low salinity level with a mean of 89 mS m–1 for ECa, which allows for rice, wheat, and a wide range of vegetables to be grown normally.  In such situations, measures such as an optimized combination of irrigation and drainage, as well as soil amendment can be taken to adjust and control the salt content.  Particularly, flattening the land with a large-scale machine was used to improve the ability of micro-topography to influence salt migration; rice and other dry, land crops were planted in rotation in combination with utilizing salt-leaching multiple times to speed up desalinization. 
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6. Molecular cloning, expression profiling and RNA interference of a vitellogenin gene from Harmonia axyridis (Coleoptera: Coccinellidae)
LIANG Chao, LIU Ting-hui, HAN Shi-peng, HE Yun-zhuan
Journal of Integrative Agriculture    2019, 18 (10): 2311-2320.   DOI: 10.1016/S2095-3119(18)62103-3
摘要76)      PDF    收藏
In this study, the Harmonia axyridis vitellogenin gene 2 (HmVg2) sequence was identified from transcriptomic databases of female adult H. axyridis and cloned into pMD18-T vector.  The HmVg2 gene was 5 460 bp in length, and formed with an open reading frame (ORF) of 5 361 nucleotides (GenBank accession no. KY794939).  The putative molecular weight of the primary HmVg2 protein was 203.459 kDa and the predicted isoelectric point (pI) was 8.59.  HmVg2 contained a signal peptide, vitellogenin N-terminal (vitellogenin-N) domain, domain of unknown function 1943 (DUF1934) domain and von Willebrand factor type D (VWD) domain.  The developmental expression profiling showed that HmVg2 was extremely highly expressed in female insects, but was expressed at lower levels in male insects.  In female insects, HmVg2 was mainly expressed in the wing and fat body.  The double-stranded RNA-HmVg2/-GFP was injected into H. axyridis, and qRT-PCR results showed that the HmVg2 gene was specifically silenced.  The eggs laid during the first five days and the hatching rate of eggs was lower than controls after dsHmVg2 injection.  This investigation demonstrated that the HmVg2 gene plays an important role in H. axyridis reproduction and enriches the function of the insect vitellogenin gene.
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7. Rhizosphere soil bacterial community composition in soybean genotypes and feedback to soil P availability
ZHOU Tao, WANG Li, DU Yong-li, LIU Ting, LI Shu-xian, GAO Yang, LIU Wei-guo, YANG Wen-yu
Journal of Integrative Agriculture    2019, 18 (10): 2230-2241.   DOI: 10.1016/S2095-3119(18)62115-X
摘要143)      PDF    收藏
Soil with low phosphorus (P) availability and organic matter contents exists in large area of southwest of China, but some soybean genotypes still show well adaptations to this low yield farmland.  However, to date, the underlying mechanisms of how soybean regulates soil P availability still remains unclear, like microbe-induced changes.  The objective of the present study was to compare the differences of rhizosphere bacterial community composition between E311 and E109 in P-sufficiency (10.2 mg kg–1) and P-insufficiency (5.5 mg kg–1), respectively, which then feedback to soil P availability.  In P-sufficiency, significant differences of the bacterial community composition were observed, with fast-growth bacterial phylum Proteobacteria, genus Dechloromonas, Pseudomonas, Massilia, and Propionibacterium that showed greater relative abundances in E311 compared to E109, while in P-insufficiency were not.  A similar result was obtained  that E311 and E109 were clustered together in P-insufficiency rather than in P-sufficiency by using principal component analysis and hierarchical clustering analysis.  The quadratic relationships between bacterial diversity and soil P availability in rhizosphere were analyzed, confirming that bacterial diversity enhanced the soil P availability.  Moreover, the high abundance of Pseudomonas and Massilia in the rhizosphere of E311 might increased the P availability.  In the present study, the soybean E311 showed capability of shaping rhizosphere bacterial diversity, and subsequently, increasing soil P availability.  This study provided a strategy for rhizosphere management through soybean genotype selection and breeding to increase P use efficiency, or upgrade middle or low yield farmland.
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8. Shade stress decreases stem strength of soybean through restraining lignin biosynthesis
LIU Wei-guo, Sajad Hussain, LIU Ting, ZOU Jun-lin, REN Meng-lu, ZHOU Tao, LIU Jiang, YANG Feng, YANG Wen-yu
Journal of Integrative Agriculture    2019, 18 (1): 43-53.   DOI: 10.1016/S2095-3119(18)61905-7
摘要286)      PDF    收藏
Lodging is the most important constraint for soybean growth at seedling stage in maize-soybean relay strip intercropping system.  In the field experiments, three soybean cultivars Nandou 032-4 (shade susceptible cultivar; B1), Jiuyuehuang (moderately shade tolerant cultivar; B2), and Nandou 12 (shade tolerant cultivar; B3) were used to evaluate the relationship between stem stress and lignin metabolism in the stem of soybean.  Results showed that the intercropped soybean was in variable light condition throughout the day time and co-growth stage with maize.  The xylem area and cross section ratio played a main role to form the stem stress.  The B3 both in intercropping and monocropping expressed a high stem stress with higher xylem area, lignin content, and activity of enzymes (phenylalanine ammonia-lyase (PAL), 4-coumarate: CoA ligase (4CL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD)) than those of B1 and B2.  Among the soybean cultivars and planting pattern, lignin content was positively correlated with stem stress.  However, a negative correlation was found between lignin content and actual rate of lodging.  In conclusion, the shade tolerant soybean cultivar had larger xylem area, higher lignin content and activities of CAD, 4CL, PAL, and POD than other soybean cultivars in intercropping.  The lodging in maize-soybean intercropping can be minimized by planting shade tolerant and lodging resistant cultivar of soybean.  The lignin content in stem could be a useful indicator for the evaluation of lodging resistance of soybean in intercropping and activities of enzymes were the key factors that influence the lignin biosynthesis.
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9. Effect of shade stress on lignin biosynthesis in soybean stems
LIU Wei-guo, REN Meng-lu, LIU Ting, DU Yong-li, ZHOU Tao, LIU Xiao-ming, LIU Jiang, Sajad Hussain, YANG Wen-yu
Journal of Integrative Agriculture    2018, 17 (07): 1594-1604.   DOI: 10.1016/S2095-3119(17)61807-0
摘要509)      PDF    收藏
To clarify how shade stress affects lignin biosynthesis in soybean stem, two varieties, Nandou 12 (shade tolerant) and Nan 032-4 (shade susceptible) grew under normal light and shade conditions (the photosynthetically active radiation and the ratio of red:far-red were lower than normal light condition).  Lignin accumulation, transcripts of genes involved in lignin biosynthesis, and intermediates content of lignin biosynthesis were analyzed.  Both soybean varieties suffered shade stress had increased plant heights and internode lengths, and reduced stem diameters and lignin accumulation in stems.  The expression levels of lignin-related genes were significantly influenced by shade stress, with interactions between the light environment and variety.  The gene of 3-hydroxylase (C3H), cinnamoyl-CoA reductase (CCR), caffeoyl-CoA O-methyltransferase (CCoAOMT), and peroxidase (POD) attributed to lignin biosynthesis under shade stress, and the down-regulation of these genes resulted in lower caffeic, sinapic, and ferulic acid levels, which caused a further decrease in lignin biosynthesis.  Under shade stress, the shade tolerant soybean variety (Nandou 12) showed stiffer stems, higher lignin content, and greater gene expression level and higher metabolite contents than shade susceptible one.  So these characteristics could be used for screening the shade-tolerant soybean for intercropping.
 
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10. Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen) 
JIA Miao, QIN Guo-hua, LIU Ting, ZHANG Jian-zhen, ZHANG Xue-yao, ZHU Kun-yan, GUO Yaping, MA En-bo
Journal of Integrative Agriculture    2011, 10 (10): 1570-1576.   DOI: 10.1016/S1671-2927(11)60153-3
摘要1822)      PDF    收藏
A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.
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