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Antifungal Activity of Poncirus trifoliata Roots against Colletotrichum Species
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作者 Kumudini M. Meepagala Tamang Prabin 《Agricultural Sciences》 CAS 2023年第3期346-355,共10页
Fruits and vegetables are an essential part of a healthy diet, providing humans with vitamins, phytonutrients, and minerals. They are significantly vulnerable, however, to post-harvest diseases caused by numerous fung... Fruits and vegetables are an essential part of a healthy diet, providing humans with vitamins, phytonutrients, and minerals. They are significantly vulnerable, however, to post-harvest diseases caused by numerous fungal and bacterial pathogens. These pathogens can cause significant quantitative and qualitative losses from harvest to consumption during the handling and storage processes. Chemical fungicides are commonly used but are likely to leave residues on the produce, rendering short shelf-life produce, such as berries, unsuitable for human consumption. Identifying eco-friendly methods to control post-harvest disease is, therefore, of utmost importance. The presence of antifungal constituents in the roots of Poncirus trifoliata extracts was detected by thin layer chromatography-based bioautography. The active constituents were isolated and identified by bioautography assay-guided fractionation using flash chromatography followed by spectroscopic techniques. In this study, xanthoxyletin, demethylsuberosin, dentatin, nordentatin, ponfolin, and clausarin were isolated from the root extracts. The antifungal activity of these compounds was moderate to weak compared to the commercial fungicide captan. This study reports the isolation and identification of natural compounds from Poncirus trifoliata that exhibited antifungal activity against Colletotrichum fragariae and Botrytis cinerea, two major post-harvest pathogens. 展开更多
关键词 poncirus trifoliata BIOAUTOGRAPHY Colletotrichum fragariae FUNGICIDE
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沙培条件下磷、钾、钙亏缺对枳(Poncirus trifoliata)幼苗根系形态及营养吸收的影响 被引量:38
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作者 曹秀 夏仁学 +2 位作者 杨环宇 张德健 赵禹 《植物营养与肥料学报》 CAS CSCD 北大核心 2014年第4期981-988,共8页
【目的】根系是吸收水分和养分的主要器官,对于多年生木本果树的生长发育尤为重要。由于果树根系庞大、分布错综复杂,对根系构型和空间分布等的研究相对较少。本文利用计算机扫描系统及其图像分析软件观测根系二维形态参数,并用原子吸... 【目的】根系是吸收水分和养分的主要器官,对于多年生木本果树的生长发育尤为重要。由于果树根系庞大、分布错综复杂,对根系构型和空间分布等的研究相对较少。本文利用计算机扫描系统及其图像分析软件观测根系二维形态参数,并用原子吸收法测定植株养分含量,以探索养分亏缺对枳根系形态的影响,以及根系形态变化与植株养分吸收的关系。【方法】本试验在沙培条件下,以柑橘砧木枳(Poncirus trifoliata)实生苗为试材,研究了缺磷、钾和钙对其根系形态以及植株体内营养元素含量的影响。利用爱普生数码扫描仪(Expression 10000XL 1.0,Epson Inc.Japan)扫描根系,并用WinRhizo Pro(S)v.2004b软件(Regent Instruments Inc.,Canada)分析获取根系总长、总体积和总表面积等二维形态参数。利用原子吸收光谱仪(SPECTR AA220)测定样品K、Ca、Mg、Fe、Cu、Zn等元素含量。【结果】从扫描数据可知,3种缺素处理对枳幼苗根系形态结构有较大的影响,即在缺素时根系总长、表面积和体积均显著降低,缺P处理使枳实生苗粗根的根系长度、表面积、体积显著降低,但是增加了中等粗根的表面积和体积;缺K和缺Ca处理的中等粗根根系长度、表面积和体积均显著低于对照;3种缺素处理均显著降低了细根和粗根的根系长度、表面积和体积。不同缺素处理对植株根系和地上部生长的影响也存在差异,缺P对地上部的抑制作用显著强于根系,缺K处理对根系生长的抑制作用较强,缺Ca对根系和地上部生长的抑制作用均较小。缺Ca处理植株体内Zn和Fe浓度均显著增加但Ca浓度降低,缺P时植株体内Ca和地上部Zn浓度增加,而缺K时植株体内Ca、Zn浓度增加但Fe和K浓度均显著降低。【结论】不同缺素条件下枳实生苗根系形态各异,导致根系对不同养分的吸收能力不同,从而使植株体内K、Ca、Zn、Fe等养分含量发生改变,最终影响整个植株根系和地上部的生长,表现出各缺素环境所特有的树体特征。 展开更多
关键词 柑橘砧木 缺素处理 根系形态参数 养分含量
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Expression of Ferric Chelate Reductase Gene in Citrus junos and Poncirus trifoliataTissues 被引量:1
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作者 李凌 范艳华 +2 位作者 罗小英 裴炎 周泽扬 《Acta Botanica Sinica》 CSCD 2002年第7期771-774,共4页
It has been hypothesized that under iron stress high ferric chelate reductase (FCR) activity in the absorptive root of plants tolerant to iron_deficiency will be induced and result in subsequent Fe 2+ transport a... It has been hypothesized that under iron stress high ferric chelate reductase (FCR) activity in the absorptive root of plants tolerant to iron_deficiency will be induced and result in subsequent Fe 2+ transport across the plasmalemma. The activity of FCR and expression of FCR gene (FRO2) in Citrus junos Sieb. ex Tanaka tolerant to iron_deficiency and Poncirus trifoliata (L.) Raf. susceptible to iron_deficiency were determined to elucidate the physiological difference which causes the different tolerance of the two citrus rootstocks to iron stress. The activity of FCR was detectable in excised roots and was stimulated about 20_times in C. junos and only about 3_times in P. trifoliata under iron deficiency for four weeks. The FRO2 of Arabidopsis was used as a probe, the tissue print technique was used to ascertain the expression of the FCR gene in C. junos and P. trifoliata under iron stress. High_level transcripts were observed in the absorptive root, young green stem as well as new leaf of C. junos under iron stress for two weeks, and the transcripts were accumulated only slightly in P. trifoliata at the same time. The results showed that the obvious increase of FCR activity was an important reason for the tolerance of C. junos to iron_deficiency, and the regulation of FCR activity seemed to be at the transcriptional level, and the expression of FRO2 occurred in the root, stem and leaf. 展开更多
关键词 Citrus junos poncirus trifoliata iron stress ferric chelate_reductase tissue print FRO2 gene Northern hybridization
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抗寒资源枳(Poncirus trifoliata)miR168a克隆、鉴定及遗传转化 被引量:2
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作者 张晓娜 刘继红 《华中农业大学学报》 CAS CSCD 北大核心 2015年第6期14-20,共7页
基于生物信息学和实验预测,克隆了柑橘抗寒资源枳ptf-miR168a成熟序列和前体序列,并预测了其靶基因。结果表明:ptf-miR168a在植物中高度保守,靶基因是NAC(UCU2)和锌指结构基因(FEZ)家族同源基因;同时,构建了ptf-miR168a前体的超表达载... 基于生物信息学和实验预测,克隆了柑橘抗寒资源枳ptf-miR168a成熟序列和前体序列,并预测了其靶基因。结果表明:ptf-miR168a在植物中高度保守,靶基因是NAC(UCU2)和锌指结构基因(FEZ)家族同源基因;同时,构建了ptf-miR168a前体的超表达载体并在烟草中进行遗传转化,得到了18个转基因系,为进一步鉴定ptf-miR168a在低温胁迫应答中的调控作用奠定基础。 展开更多
关键词 miR168 抗寒性 靶基因 克隆 遗传转化 低温胁迫
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The extract of the immature fruit of Poncirus trifoliata induces apoptosis in colorectal cancer cells via mitochondrial autophagy 被引量:1
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作者 Sun-Young Kim Ho-Keun Yi +4 位作者 Bong-Sik Yun Dae-Yeol Lee Pyung Han Hwang Hae-Ryong Park Min Sun Kim 《Food Science and Human Wellness》 SCIE 2020年第3期237-244,共8页
The immature fruits of Poncirus trifoliate are used as a medicine for the treatment of gastrointestinal disorders,inflammation,and allergies in East Asia.However,their effect on colon cancer cells remains unclear.We i... The immature fruits of Poncirus trifoliate are used as a medicine for the treatment of gastrointestinal disorders,inflammation,and allergies in East Asia.However,their effect on colon cancer cells remains unclear.We investigated the effect of the immature fruit of P.trifoliate extract on colorectal adenocarcinoma.The extract of the immature fruit of P.trifoliata inhibited the proliferation of CT-26 cells compared with untreated cells and it induced autophagy and apoptosis through the protein kinase B/mammalian target of rapamycin and 5'-AMP-activated protein kinase pathways.The number of autophagic vacuoles and autophage markers increased in response to the extract.At later time-points,apoptosis increased dose/time-dependently.In CT-26 cells pre-treated a pan-caspase inhibitor prior to P.trifoliata immature fruit extract treatment,we did not find any change in pro-caspase 3 and pro-PARP levels.Additionally,in cells pre-treated autphage inhibitor,SQSTM1/p62 and LC3AB,pro-caspase 3 and pro-PARP levels did not change.Our results indicate the molecular mechanisms that the extract of the immature fruit of P.trifoliata induces apoptosis in colorectal carcinoma cells by inducing mitochondrial autophagy.In this study,we provided a draft for further investigate the use of MEPT for colorectal cancer inhibition. 展开更多
关键词 ANTICANCER AUTOPHAGY APOPTOSIS MITOCHONDRIA poncirus trifoliata
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DR5 is a Suitable System for Studying the Auxin Response in the Poncirus trifoliata-Xanthomonas axonopodis pv.citri Interaction 被引量:1
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作者 Xuan Xiao Xiaoyan Li +1 位作者 Chunli Chen Wenwu Guo 《Horticultural Plant Journal》 SCIE 2020年第5期277-283,共7页
The local auxin distribution characteristics in the roots,stems,and leaves of stably transformed plantlets of trifoliate orange(Poncirus trifoliata)with auxin reporter system DR5::GUS-YFP were elucidated in this resea... The local auxin distribution characteristics in the roots,stems,and leaves of stably transformed plantlets of trifoliate orange(Poncirus trifoliata)with auxin reporter system DR5::GUS-YFP were elucidated in this research.The auxin response maxima could be observed in the apex of the root tip,primary phloem of the tender stem,and the margin of the young leaves according to the activity of theβ-glucuronidase(GUS)reporter gene triggered by the auxin responsive DR5 promoter.Auxin responses in the apex of the root tips increased when treated with synthetic auxin 1-naphthylacetic acid(NAA),but decreased when treated with the auxin polar transportation inhibitor 2,3,5-triiodobenzoic acid(TIBA).These results indicated that the DR5 reporter system worked in P.trifoliata for auxin distribution and response observation.Trifoliate orange is highly susceptible to citrus canker disease.Auxin accumulation was observed visually in the invasion sites of the detached leaves inoculated with Xanthomonas axonopodis pv.citri(Xac)by GUS staining;the upregulated expression of the YFP,GH3.1,GH3.9,and SAUR genes assessed by quantitative real-time PCR(qRT-PCR)also identified auxin accumulation in the inoculated tissues following Xac infection.Overall,these findings indicated that the plantlets of P.trifoliata engineered with the auxin reporter gene provided a promising system for studying auxin responses during Xac infection. 展开更多
关键词 poncirus trifoliata DR5 AUXIN Xanthomonas axonopodis pv.citri
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Extraction and characterization of gelling pectin from the peel of Poncirus trifoliata fruit 被引量:1
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作者 Kouassi L.Koffi Beda M.Yapo V.Besson 《Agricultural Sciences》 2013年第11期614-619,共6页
In the framework of searching for new pectin sources to partially compensate for domestic and regional demands, the peel (albedo) of the “non-comestible” fruit of Poncirus trifoliata was investigated using a relativ... In the framework of searching for new pectin sources to partially compensate for domestic and regional demands, the peel (albedo) of the “non-comestible” fruit of Poncirus trifoliata was investigated using a relatively simple experimental design for optimization, in which only the variable was the extraction pH (1.0, 1.5, and 2.0) on the basis of our previous studies on diverse pectin sources. The results showed that the yield of pectin (7.4%-19.8%) was strongly influenced by the extraction pH when the other parameters, namely the solid to liquid extractant (S/L) ratio, temperature (T &degC), and time (t) were fixed to 1:25 (w/v), 75&degC, and 90 min, respectively. Likewise, the galacturonic acid content (GalA: 61.4%-79.2%), total neutral sugar content (TNS: 9.1%-22.5%), degree of branching (3.5%-13.9%), homogalacturonan (HG) to rhamnogalacturonan-I (RG-I) ratio (2.2-5.6), degree of methylesterification (DM: 54-77), viscosity average molecular weight (Mν: 57-82), and gelling capacity (GC: 124-158) were all affected by the extraction pH. The optimum pH for producing pectin with good yield, quality characteristics (GalA > 65%, DM > 60, Mν > 80 kDa), and gelling capacity (GC > 150), from the peel of P. trifoliata fruit, was found to be pH 1.5. 展开更多
关键词 poncirus trifoliata PECTIN Block Copolymers Macromolecular Characteristics Gelling Strength
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枳PtMLP1启动子的克隆和表达分析
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作者 姚利晓 苏娟 +4 位作者 郭兴茹 李凤龙 何永睿 邹修平 陈善春 《中国农业科学》 CSCD 北大核心 2023年第24期4906-4915,共10页
【目的】基因工程是柑橘品种改良的一种重要手段。本研究基于枳根消减文库中主要乳胶蛋白基因PtMLP1片段,克隆根特异启动子序列,为研究外源基因在柑橘根组织的特异表达奠定基础。【方法】同源克隆PtMLP1及启动子序列。利用ExPASy、PSIP... 【目的】基因工程是柑橘品种改良的一种重要手段。本研究基于枳根消减文库中主要乳胶蛋白基因PtMLP1片段,克隆根特异启动子序列,为研究外源基因在柑橘根组织的特异表达奠定基础。【方法】同源克隆PtMLP1及启动子序列。利用ExPASy、PSIPRED、SWISS-MODEL等在线软件对PtMLP1编码蛋白的理化特征、二级结构和三级结构进行生物信息学分析,利用PlantCARE数据库对PtMLP1启动子的顺式作用元件进行预测。实时荧光定量PCR法对PtMLP1在不同树龄枳根和叶中的表达进行分析。构建PtMLP1启动子与GUS标记基因的融合载体,利用根癌农杆菌转化法转化枳上胚轴,GUS染色观察标记基因的表达部位。【结果】枳PtMLP1含2个外显子和1个内含子,开放阅读框长471 bp。PtMLP1蛋白由156个氨基酸组成,分子量17.63 kDa,等电点5.49,含Bet v I功能域。其二级结构含3个α-螺旋和7个β-折叠,三级结构包含一个保守疏水基结合位点和一个富含甘氨酸的回环结构。5′端1666 bp的上游调控序列不仅有TATA-box、CAAT-box等启动子结构的核心元件,还具有多个根组织特异表达元件,以及TGACG-motif、P-box和ABRE等激素应答相关的顺式作用元件。3′端非翻译区具有加尾信号AATAAA。该基因在1月龄苗、6月龄苗、20年生成年枳根中的表达量分别是叶中的46.34、74.82、110.25倍。构建启动子的融合表达载体pBI121-ProPtMLP1::GUS,获得枳转基因植株。PtMLP1启动子驱动GUS在转基因枳幼苗根中特异表达,GUS在3个转基因枳株系的根中表达量分别为叶中表达量的124.78、11.53和7.76倍。【结论】获得柑橘主要乳胶蛋白PtMLP1及启动子序列,该启动子可驱动标记基因在柑橘根组织特异表达。 展开更多
关键词 主要乳胶蛋白 根特异性启动子 GUS
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枳LEA基因家族鉴定及其对非生物胁迫的响应 被引量:2
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作者 陈凯 佟晓楠 +3 位作者 张晓媛 范丽珍 钟静 李兴涛 《西北植物学报》 CAS CSCD 北大核心 2023年第6期918-928,共11页
为明确LEA基因在枳非生物胁迫中的作用,研究通过生物信息学方法对枳LEA基因家族进行全基因组的鉴定,并采用qRT-PCR技术分析其在不同非生物胁迫下的表达情况。结果表明:(1)枳基因组中鉴定得到57个LEA基因家族成员,编码蛋白的氨基酸长度在... 为明确LEA基因在枳非生物胁迫中的作用,研究通过生物信息学方法对枳LEA基因家族进行全基因组的鉴定,并采用qRT-PCR技术分析其在不同非生物胁迫下的表达情况。结果表明:(1)枳基因组中鉴定得到57个LEA基因家族成员,编码蛋白的氨基酸长度在62~945 aa之间,分子质量在6.95~104.98 kD之间。(2)系统进化分析表明,PtrLEA基因可分为8个亚家族,LEA_1~LEA_6、dehydrin和SMP,LEA_2亚族家族成员最多。(3)顺式作用元件分析表明,PtrLEA启动子上存在大量的植物激素、胁迫响应以及生长发育有关的响应元件。(4)转录组数据分析结果显示,LEA基因家族具有组织表达特异性,PtrLEA 15、PtrLEA 17、PtrLEA 23、PtrLEA 51在所有组织中均有高表达,也发现有3个基因在所有组织中不表达。(5)实时荧光定量PCR结果显示:PtrLEA基因在低温、干旱和高盐胁迫处理条件下,与对照相比分别有6、2和6个基因上调表达,推测该基因家族参与多种非生物胁迫应答。 展开更多
关键词 lEA基因家族 逆境响应 表达模式
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枳SPL9和SPL13全长cDNA克隆、亚细胞定位和表达分析 被引量:18
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作者 宋长年 钱剑林 +4 位作者 房经贵 王化坤 邱学林 章镇 张晓莹 《中国农业科学》 CAS CSCD 北大核心 2010年第10期2105-2114,共10页
【目的】从枳[Poncirus trifoliata(L.)Raf.]中克隆SBP类[SQUAMOSA(SQUA)promoter-binding-like]转录因子基因SPL9和SPL13全长,构建SPL9和SPL13亚细胞定位表达载体验证其是否具有核定位功能,利用荧光定量PCR研究其在枳不同组织的表达特... 【目的】从枳[Poncirus trifoliata(L.)Raf.]中克隆SBP类[SQUAMOSA(SQUA)promoter-binding-like]转录因子基因SPL9和SPL13全长,构建SPL9和SPL13亚细胞定位表达载体验证其是否具有核定位功能,利用荧光定量PCR研究其在枳不同组织的表达特性,初步确定SPL9和SPL13在枳生长发育过程中的作用。【方法】利用生物信息学结合RACE技术以枳花器官的cDNA为模板,克隆出SPL9和SPL13基因全长,分别命名Pt-SPL9和Pt-SPL13,大小分别是1519bp和1824bp,在GenBank的登录号分别是FJ502237和FJ502238;构建Pt-SPL9和Pt-SPL13亚细胞定位载体35S-GW-FJ502237/FJ502238-GFP,基因枪转化洋葱表皮细胞,暗培养24h后激光共聚焦显微镜下观察;利用SYBR Green I实时定量RT-PCR方法检测Pt-SPL9和Pt-SPL13在根、茎、叶、花序、花和果等不同组织中的表达。【结果】生物信息学分析表明,Pt-SPL9和Pt-SPL13的cDNA序列中都有microRNA156的识别位点,Pt-SPL9与金鱼草、拟南芥和玉米SPL9的同源性分别为48.9%、42.5%和41.7%;Pt-SPL13与拟南芥SPL13、水稻的SPL16和玉米的TGA1同源性分别为40.8%、38.1%和35.8%。Pt-SPL9和Pt-SPL13与其它植物的SBP一样有着高度保守的序列,即SBP结构域和一个双向核定位信号KRXXXRRRK。亚细胞定位结果表明,Pt-SPL9和Pt-SPL13均定位于细胞核中。SYBR Green I实时定量RT-PCR结果表明,Pt-SPL9和Pt-SPL13在各个器官均有表达,但表达量不同,Pt-SPL9在茎中的表达量最高,在花和叶中的表达量次之,在根、花芽和幼果中的表达量最低;Pt-SPL13在幼果中的表达量最高,在茎和花芽中的表达量相当,其次为叶,在花和根中的表达量很低。【结论】转录因子Pt-SPL9和Pt-SPL13均具有核定位功能,Pt-SPL9和Pt-SPL13对枳的茎和果实的发育可能有着重要作用。 展开更多
关键词 SPl9和SPl13 亚细胞定位 荧光定量RT-PCR
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枳NLP转录因子克隆及其在不同水分条件下的表达 被引量:6
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作者 曹雄军 卢晓鹏 +4 位作者 熊江 李静 吴倩 周芳芳 谢深喜 《中国农业科学》 CAS CSCD 北大核心 2016年第2期381-390,共10页
【目的】分析柑橘砧木枳(Poncirus trifoliata(L.)Raf.)参与氮素吸收与同化的重要转录因子NLP在干旱胁迫下的表达模式,为柑橘在干旱胁迫条件下对氮素吸收与同化的调控机理提供研究参考。【方法】以柑橘砧木枳为试验材料,在甜橙(Citrus s... 【目的】分析柑橘砧木枳(Poncirus trifoliata(L.)Raf.)参与氮素吸收与同化的重要转录因子NLP在干旱胁迫下的表达模式,为柑橘在干旱胁迫条件下对氮素吸收与同化的调控机理提供研究参考。【方法】以柑橘砧木枳为试验材料,在甜橙(Citrus sinensis(L.)Osb.)基因组数据库的基础上,利用生物信息学筛选获得甜橙NLP,并以此设计引物扩增枳NLP的CDS全长并测序。利用Clustal X和MEGA程序进行序列比对和系统发育分析,利用实时荧光定量PCR技术分析不同水分条件下的枳NLP的表达模式。【结果】筛选并克隆获得了4条枳NLP序列,为Pt NLP2、Pt NLP4、Pt NLP7和Pt NLP8。序列比对分析表明枳NLP之间的蛋白质序列一致性为45.13%,均具有明显的RWP-RK和PB1结构域。枳与甜橙的NLP蛋白质序列的一致性极高,分别为97.57%、96.47%、99.0%及97.33%。系统发育分析表明枳NLP可分为4个进化类型,与拟南芥的NLP分类一致,即Pt NLP2与At NLP1/2一类,Pt NLP4与At NLP4/5一类,Pt NLP7与At NLP6/7一类,Pt NLP8与At NLP8/9一类。在不同水分条件下,枳NLP的表达模式在叶与根中存在差异。随着基质水分含量的减少,枳叶NLP呈现先上升再下降的趋势。与对照(相对持水量为61.0%)相比,叶Pt NLP2、Pt NLP4、Pt NLP7及Pt NLP8的表达量在相对持水量为15.4%时上调至最高水平,分别上调了2.9、3.5、5.9及2.8倍,之后持续下调;到相对持水量为9.4%时,其表达水平低于对照但无显著差异。而枳根NLP的表达量在对照中最高,后随着水分的散失呈总体下降趋势且差异显著,与对照相比,根Pt NLP2、Pt NLP4、Pt NLP7及Pt NLP8的最大下调倍数分别为6.7、2.8、4.8及2.3。复水后,枳叶与根NLP的表达量低于复水前,且与对照差异显著。【结论】枳NLP的表达与土壤的水分条件密切相关。枳叶片NLP的表达水平在干旱胁迫前、中期上调,后期下调;而枳根NLP的表达水平在干旱胁迫下持续下调。其中,Pt NLP2和Pt NLP7的表达量在枳根响应干旱胁迫中变化较大。 展开更多
关键词 NlP转录因子 干旱胁迫 基因表达
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光照、温度和NaCl胁迫对枳壳种子萌发及幼苗生长的影响 被引量:2
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作者 王贵元 李艳 《长江大学学报(自然科学版)》 CAS 2011年第7期237-240,285,共4页
采集未成熟的枳壳[Poncirus trifoliate(L.)Raf.]果实剥取幼嫩种子播种,研究了枳壳种子的萌发情况和幼苗的生长状况。试验设置12个处理,分别为:(1)光照处理:设全光照、光照∶黑暗=12h∶12h、全黑暗3个处理;(2)温度处理:在光照∶黑暗=12h... 采集未成熟的枳壳[Poncirus trifoliate(L.)Raf.]果实剥取幼嫩种子播种,研究了枳壳种子的萌发情况和幼苗的生长状况。试验设置12个处理,分别为:(1)光照处理:设全光照、光照∶黑暗=12h∶12h、全黑暗3个处理;(2)温度处理:在光照∶黑暗=12h∶12h,设15、25、35℃3个恒温和25℃/15℃(昼夜交替)1个变温,共4个处理;(3)氯化钠(NaCl)胁迫处理:在光照∶黑暗=12h∶12h和25℃条件下,NaCl设置0.0(CK)、0.1、0.3、0.5、0.7、0.9mol/L共6个处理。结果表明,枳壳种子萌发和幼苗生长的最佳处理组合均为光照∶黑暗=12h∶12h、25℃和0.0mol/L的NaCl。 展开更多
关键词 枳壳[poncirus trifoliate(l.)Raf.] 种子 发芽率 发芽势 幼苗 光照 温度 氯化钠
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CclSAUR49基因的表达特征及对类柠檬苦素生物合成的影响 被引量:1
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作者 余洪 易倩 +3 位作者 张曼曼 朱世平 王福生 赵晓春 《果树学报》 CAS CSCD 北大核心 2021年第8期1240-1251,共12页
【目的】探究CclSAUR49基因对柑橘类柠檬苦素生物合成的影响。【方法】通过高效液相色谱(HPLC)检测叶片中的诺米林和柠檬苦素含量;利用实时荧光定量PCR分析基因的表达量。分别从柚和湖北早实枳中克隆CclSAUR49基因编码序列(coding seque... 【目的】探究CclSAUR49基因对柑橘类柠檬苦素生物合成的影响。【方法】通过高效液相色谱(HPLC)检测叶片中的诺米林和柠檬苦素含量;利用实时荧光定量PCR分析基因的表达量。分别从柚和湖北早实枳中克隆CclSAUR49基因编码序列(coding sequence,CDS)及启动子,利用瞬时转化烟草对CclSAUR49蛋白进行表达定位;通过遗传转化湖北早实枳分析该基因的组织表达特性及其对类柠檬苦素生物合成的影响。【结果】类柠檬苦素含量和CclSAUR49表达水平在2个沙田柚品种间以及叶片的生长发育中均呈显著负相关。CclSUAR49蛋白定位于质膜和细胞核。从湖北早实枳中克隆得到的CclSAUR49基因的启动子序列中不含吲哚乙酸(IAA)响应元件。启动子活性分析发现,Ccl-SAUR49基因主要在茎、根和老叶中表达,在嫩叶中没有表达。超量表达CclSAUR49导致类柠檬苦素含量显著降低,植株的叶片和节间显著伸长,茎增粗。【结论】柑橘CclSAUR49基因的表达有明显的组织特异性,对类柠檬苦素的合成或积累有抑制作用。 展开更多
关键词 CclSAUR49基因 类柠檬苦素 启动子 顺式作用元件 超量表达
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枳NLP转录因子响应干旱胁迫并与NRE顺式作用元件互作 被引量:3
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作者 曹雄军 卢晓鹏 +2 位作者 熊江 李静 谢深喜 《中国农业科学》 CAS CSCD 北大核心 2018年第17期3370-3378,共9页
【目的】探讨枳氮素含量在干旱条件下的变化,以及氮代谢途径枳亚硝酸还原酶基因(NiR)和NLP转录因子的响应表达,分析确定枳NLP转录因子与NiR启动子区域硝酸盐响应顺式作用元件(nitrate-responsive cis-element,NRE)位点绑定互作。【方法... 【目的】探讨枳氮素含量在干旱条件下的变化,以及氮代谢途径枳亚硝酸还原酶基因(NiR)和NLP转录因子的响应表达,分析确定枳NLP转录因子与NiR启动子区域硝酸盐响应顺式作用元件(nitrate-responsive cis-element,NRE)位点绑定互作。【方法】以一年生的砧木枳为试验材料,进行干旱处理直至植株叶片开始萎蔫,并设置对照。利用凯氏定氮仪检测干旱条件下枳叶片和侧根的全氮含量,并同步利用实时荧光定量PCR(RT-qPCR)和2-ΔΔCT法分析PtNiR、PtNLP2、PtNLP4、PtNLP7及PtNLP8在枳叶片和侧根的表达情况。通过基因克隆并检索与比对分析,获取PtNiR启动子区域的硝酸盐响应顺式作用元件序列,之后利用同源克隆方法构建用于酵母单杂交的pHIS2-4×NRE和p GADT7-Rec-NLP载体,将构建的pHIS2-4×NRE和p GADT7-Rec-NLP载体质粒共同转化到酵母菌株Y187中,在氨基酸缺失培养基(SD/-Trp/-Leu,以及SD/-His/-Trp/-Leu/+120 mmol·L^(-1) 3-氨基三唑)上30℃恒温培养3—5 d,观察酵母的生长情况,以确定枳NLP转录因子(PtNLP2、PtNLP4、PtNLP7及PtNLP8)与NRE的互作关系。【结果】受干旱的影响,枳叶片全氮含量表现出先升高再下降的趋势,而侧根中全氮含量则显著下降;在对照中,枳叶片和侧根全氮含量均显著上升。RT-qPCR分析表明,PtNiR的表达水平在叶片中先上调然后再显著下调,而在侧根中则随着干旱程度的增加显著下调;PtNLP2、PtNLP4及PtNLP7在枳叶片和侧根中的表达也均呈现不同程度的先上调然后再受到抑制的规律,而PtNLP8在枳叶片和侧根中的表达则在一定程度上受到干旱抑制。通过克隆测序分析,在枳NiR启动子区域-196至-154的位置发现了疑似NRE。酵母单杂交分析表明,转化pHIS2-4×NRE-HIS3载体和p GADT7-Rec-NLP载体的Y187酵母在对照培养基(-Trp/-Leu)和含有120 mmol·L^(-1) 3-氨基三唑的三缺培养基(-His/-Trp/-Leu)上均能正常生长。【结论】枳侧根中的全氮含量受干旱的影响而显著下降,以对照为参比,枳侧根和叶片中的全氮含量均随着干旱时间的延长而相对减少。氮代谢途径的枳NiR和NLP转录因子在叶片和侧根中的表达对干旱有不同程度的响应。PtNLP2、PtNLP4、PtNLP7及PtNLP8转录因子均能与PtNiR启动子中的硝酸盐响应顺式作用元件(NRE)位点绑定互作,从而激活下游基因的表达。 展开更多
关键词 NlP转录因子 干旱胁迫 硝酸盐响应顺式作用元件 绑定
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Marker Assisted Selection in Citrus Rootstock Breeding Based on a Major Gene Locus ‘Tyr1’Controlling Citrus Nematode Resistance
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作者 XIANG Xu DENG Zhan-ao +2 位作者 CHEN Chun-xian Fred G Gmitter Jr Kim Bowman 《Agricultural Sciences in China》 CAS CSCD 2010年第4期557-567,共11页
Based on the former constructed 'Tyrl' locus genetic map in family 9145, from LB6-2 [Clementine mandarin (C. reticulata) × Hamlin orange (C. sinensis)] × Swingle citrumelo (C. paradise × P. trifo... Based on the former constructed 'Tyrl' locus genetic map in family 9145, from LB6-2 [Clementine mandarin (C. reticulata) × Hamlin orange (C. sinensis)] × Swingle citrumelo (C. paradise × P. trifoliata), 9 markers were chosen for application in evaluating their effectiveness in marker-assisted selection (MAS) for citrus rootstock breeding program from many F1 progeny of Poncirus trifoliata. As the mapping revealed that these markers were estimated within a range of 12.1 cM in the linkage group, and among them, SCO07 co-segregated with "Tyrl', and 7A4R as the closest to 'Tyrl' with a distance of 1.5 cM, these markers were basically fitful to go MAS screening. The results of screening P. trifoliata F1 progeny indicated that all the markers were inherited in codominant fashion and most of them were heterozygous on PT (Pomery of P. trifoliata)., marker 4L17R/CfoI and 7A4(1407)/BfaI were proved to be consistently reliable for accurate scoring of genotypes and the revealed polymorphism was basically coincided with the citrus nematode resistant phenotype within tested populations. The polymorphic genotype with marker 4L17R/Cfol was found completely matched up with the phenotype of individuals that conferred high resistance to citrus nematode when the USDA hybrid rootstocks were screened. Utilization of these markers, especially the highly specific 4L17R/Cfol and 7A4(1407)/Bfal, should result in great benefit to world citrus industry for early selection in rootstock-breeding program. 展开更多
关键词 CITRUS poncirus trifoliata nematode resistance marker-assisted selection
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枳橙砧沃柑在桂南地区的综合表现 被引量:2
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作者 刘要鑫 仇惠君 +7 位作者 欧智涛 陈香玲 高兴 陈东奎 李果果 赵洪涛 黄宏明 李善彪 《南方农业学报》 CAS CSCD 北大核心 2023年第1期209-216,共8页
【目的】探究枳橙砧木对沃柑树体及果实品质的影响,为筛选桂南地区沃柑适宜的砧木提供理论依据。【方法】在广西南宁市武鸣区采取长期定位试验,对嫁接在枳橙上的沃柑果实品质进行持续3年(2018—2020年)的研究,以资阳香橙砧、枳砧和红橘... 【目的】探究枳橙砧木对沃柑树体及果实品质的影响,为筛选桂南地区沃柑适宜的砧木提供理论依据。【方法】在广西南宁市武鸣区采取长期定位试验,对嫁接在枳橙上的沃柑果实品质进行持续3年(2018—2020年)的研究,以资阳香橙砧、枳砧和红橘砧沃柑为对照,分析其树体指标(砧木干周、接穗干周、干周差、株高、挂果数量)及果实品质指标(单果重、果实横径、纵径、果形指数、果皮厚度、可食率、维生素C、可溶性固形物、柠檬酸、总糖、固酸比)等的变化。【结果】综合3年数据可知,枳橙砧沃柑在单果重、挂果数量和部分品质指标上优于其他砧木,3年平均挂果数为299个,单果重为143.07 g,维生素C含量为19.92 mg/100 m L,可溶性固形物含量为12.51%,柠檬酸含量为0.584%,总糖含量为12.46%,固酸比为21.42。枳橙砧与枳砧相比,呈现出生长势强,挂果数量多等优势,但与香橙砧相比,差异不显著(P>0.05,下同);结合单果重表现,枳橙砧沃柑无明显的下降趋势,与香橙砧沃柑相比,均能持续稳产。枳橙砧沃柑的果实品质整体表现低于枳砧沃柑,但进入丰产期后,二者品质差异不显著。对比其他3种砧木,红橘砧沃柑整体表现较差,主要为挂果数量较少,果实品质指标不理想,其中3年平均挂果数量为185个。【结论】枳橙砧沃柑在桂南地区综合表现较好,可考虑枳橙砧作为桂南地区沃柑的适宜砧木;但枳橙砧不适宜在缺锌较严重的地块种植,且在生产中,枳橙砧应控制树势,适时修剪,不适宜密植。 展开更多
关键词 枳橙 沃柑 果实品质 影响 桂南地区
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<i>Ralstonia solanacearum</i>Induction Causes Biochemical and Oxidative Stress Isozyme Variations in Mangroves without Wilt
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作者 Sasidharan Sreedevi Sreedharan Sajith +1 位作者 Kulangara Nanu Remani Sailas Benjamin 《American Journal of Plant Sciences》 2013年第10期1968-1975,共8页
We evaluated the effects of Ralstonia solanacearum (Rs) induced biotic stress in three mangroves, viz., Avicennia officinalis, Derris trifoliata and Excoecaria agallocha. These plants were grown in pots as well as hyd... We evaluated the effects of Ralstonia solanacearum (Rs) induced biotic stress in three mangroves, viz., Avicennia officinalis, Derris trifoliata and Excoecaria agallocha. These plants were grown in pots as well as hydroponic systems with sufficient controls, and about 8 × 104 colony forming units of Rs suspension was injected into the healthy test plants (saplings). The plants were subjected to biochemical and isozyme analyses. Upon induction of Rs stress, highly significant (p 0.01) biochemical changes (%) were noticed in respect to controls: carbohydrate content was generally high (24-36) in all plants;hydroponic mangroves showed higher starch content, mangroves under hydroponic system showed increased reducing sugars (29-46), almost all mangroves showed increased protein content;phenolics showed a swing of decrease or increase between plants grown in pot and hydroponic systems;and all plants in general showed higher proline content. Regarding oxidative stress isozymes (OSE) and superoxide dismutase (EC1.15.1.1), mangroves showed 1 or 2 additional isozymes with comparable relative mobility;similarly 1 or 2 additional peroxidase (EC1.11.1.7) isozymes were found in mangroves grown under hydroponic system. Briefly, Rs induced biotic stress did not cause any wilt symptom in all the 3 mangroves tested, but their normal biochemical and OSE patterns, especially of those grown as hydroponics were elicited to significantly higher levels. 展开更多
关键词 R. SOlANACEARUM AVICENNIA officinalis l. DERRIS trifoliata lour. Excoecaria agallocha l.
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枸橘药材HPLC特征指纹图谱研究
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作者 付妍 张亚双 +5 位作者 王亚琼 张华锋 陈卫 周坚 邢以文 薛满 《中国合理用药探索》 CAS 2020年第10期89-92,共4页
目的:建立枸橘药材的HPLC特征指纹图谱分析方法,为枸橘药材的真伪鉴别及质量标准的建立提供依据。方法:采用Diamonsil Plus C18-A色谱柱(250 mm×4.6 mm,5μm),以甲醇(A)-1%冰醋酸溶液(B)为流动相进行梯度洗脱,检测波长283 nm,柱温3... 目的:建立枸橘药材的HPLC特征指纹图谱分析方法,为枸橘药材的真伪鉴别及质量标准的建立提供依据。方法:采用Diamonsil Plus C18-A色谱柱(250 mm×4.6 mm,5μm),以甲醇(A)-1%冰醋酸溶液(B)为流动相进行梯度洗脱,检测波长283 nm,柱温30℃。结果:建立了枸橘药材的HPLC特征指纹图谱,标记了6个特征峰。结论:该方法简便易行,为枸橘药材的真伪鉴别提供了科学依据,为枸橘药材的质量监管提供了有力支撑。 展开更多
关键词 枸橘 HPlC 特征指纹图谱 真伪鉴别 质量标准
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睡菜水提液对小鼠的急性毒性实验研究
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作者 魏雪娇 石松云 +7 位作者 毛楚涵 何婷 刘呈雄 陈剑锋 贺海波 黄文峰 邹坤 程凡 《三峡大学学报(自然科学版)》 CAS 2023年第3期108-112,共5页
目的:观察睡菜水提液对大鼠的急性毒性.方法:预实验:灌胃给予小鼠水提液剂量分别为2455、1841.25、1380.94、1035.70 g/L,观察给药当天及7 d内小鼠的状态,计算半数致死量(50%lethal dose,LD_(50));最大耐受量实验:灌胃给予睡菜最大剂量2... 目的:观察睡菜水提液对大鼠的急性毒性.方法:预实验:灌胃给予小鼠水提液剂量分别为2455、1841.25、1380.94、1035.70 g/L,观察给药当天及7 d内小鼠的状态,计算半数致死量(50%lethal dose,LD_(50));最大耐受量实验:灌胃给予睡菜最大剂量2455 g/L,观察给药后14 d内小鼠状态和死亡情况,给药14 d后检测小鼠的血液生化指标、计算主要脏器指数以及观察主要脏器病理学变化.结果:未能测出睡菜水提液的半数致死量.最大耐受量实验显示:给药后14 d内小鼠状态良好,无中毒反应或死亡情况.给药组小鼠第3、7、11、14 d体质量增长与空白组比较无统计学差异(P>0.05);给药组小鼠血液生化指标、主要脏器指数与空白组无统计学差异(P>0.05);组织形态学、组织病理学检查未发现睡菜水提液对小鼠主要脏器产生病理改变.结论:睡菜水提液的最大无毒剂量大于2455 g/L,是成人临床用药(15 g)的100.6倍,是安全的用药剂量范围. 展开更多
关键词 睡菜 小鼠 急性毒性实验
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Common mycorrhizal networks activate salicylic acid defense responses of trifoliate orange(Poncirus trifoliata) 被引量:5
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作者 Yi-Can Zhang Ying-Ning Zou +1 位作者 Li-Ping Liu Qiang-Sheng Wu 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2019年第10期1099-1111,共13页
Citrus canker, caused by Xanthomonas axonopo-dis pv. citri ('Xac'), is an important quarantine disease in citrus crops. Arbuscular mycorrhizal fungi (AMF) form symbiotic interactions with host plants and furth... Citrus canker, caused by Xanthomonas axonopo-dis pv. citri ('Xac'), is an important quarantine disease in citrus crops. Arbuscular mycorrhizal fungi (AMF) form symbiotic interactions with host plants and further affect their disease resistance, possibly by modulating the activity of salicylic acid (SA), a key phytohormone in disease resistance. Common mycorrhizal networks (CMNs) can interconnect plants, but it is not yet clear whether CMNs promote resistance to citrus canker and, if so, whether SA signaling is involved in this process. To test this possibility, we used a two-chambered rootbox to establish CMNs between trifoliate orange (Poncirus trifoliata) seedlings in chambers inoculated (treated) or not (neighboring) with the AMF, Paraglomus occultum. A subset of the AMF-inoculated seedlings were also inoculated with Xac (+AMF+Xac). At 2 d post-inoculation (dpi), compared with the +AMF-Xac treatment, neighboring seedlings in +AMF+Xac treatment had lower expression levels of the SA biosynthetic genes, PtPAL, PtEPS1,&nbsp;and PtPBS3, but higher SA levels, which attributed to the upregulation of PtPAL and PtPBS3 in treated seedlings and the transfer of SA, via CMNs, to the neighboring seedlings. At 4 dpi, the pathogenesis-related (PR) protein genes, PtPR1, PtPR4, and PtPR5, and the transcriptional regulatory factor gene, PtNPR1, were activated in neighboring seedlings of+AMF+Xac treatment. At 9 dpi, root phenylalanine ammo-nia-lyase activity and total soluble phenol and lignin concentrations increased in neighboring seedlings of+AMF+Xac treatment, likely due to the linkage and signal transfer, via CMNs. These findings support the hypothesis that CMNs transfer the SA signal from infected to neighboring healthy seedlings, to activate defense responses and affording protection to neighboring plants against citrus canker infection. 展开更多
关键词 poncirus trifoliata MYCORRHIZAl salicylic acid DEFENSE RESPONSES
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