CN109402171A - 一种松材线虫RNAi调控基因及其应用 - Google Patents
一种松材线虫RNAi调控基因及其应用 Download PDFInfo
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- CN109402171A CN109402171A CN201811624288.4A CN201811624288A CN109402171A CN 109402171 A CN109402171 A CN 109402171A CN 201811624288 A CN201811624288 A CN 201811624288A CN 109402171 A CN109402171 A CN 109402171A
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- bursaphelenchus xylophilus
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Abstract
本发明提供了一种松材线虫RNAi调控基因，该RNAi调控基因的全长序列如SEQ ID NO:1所示，该RNAi调控基因的cDNA全长序列如SEQ ID NO:2所示。本发明还提供了所述的松材线虫RNAi调控基因在制备松材线虫防治菌剂中的应用。本发明筛选到一段高效的RNAi调控基因片段，将含有该基因片段的阳性真菌转化子饲喂松材线虫，获得明显的RNAi表型，松材线虫Bx‑etr‑1基因表达明显下调，松材线虫出现胚胎发育不正常，幼虫死亡率高，运动轨迹改变，运动速度变慢，种群数量明显下降的现象。
Description
技术领域
本发明属于生物工程的技术领域，具体涉及一种松材线虫RNAi调控基 因及其应用。
背景技术
松材线虫(Bursaphelenchus xylophilus)在松树内取食危害，引起的松材 线虫病导致松树大量地成片死亡，而且，其发病速度快，防控难度大，至今 没有很有效的可持续的防控措施。
RNA interference(RNAi)主要是指外源或内源的双链RNA(dsRNA)可 以引起与之对应的靶基因mRNA降低的现象。利用RNAi技术在植物寄生性 线虫的防控中已得到应用，即通过寄主植物介导RNAi控制农业上重要的根 结线虫和胞囊线虫的种群数量，已成为具有广泛应用前景的新的防控途径。 在松材线虫的研究中，真菌介导RNAi技术已建立。
因此，筛选高效的RNAi靶标是进一步开发松材线虫RNAi防控技术的 关键。
发明内容
为了解决上述技术问题，本发明提供了一种松材线虫RNAi调控基因及 其应用。
具体的，本发明提供了一种松材线虫RNAi调控基因，该RNAi调控基 因的全长序列如SEQ ID NO:1所示。
需要进一步说明的，该RNAi调控基因的cDNA全长序列如SEQ ID NO:2 所示。
需要进一步说明的，该RNAi调控基因的编码氨基酸序列如SEQ ID NO:3所示。
本发明还提供了一种重组表达质粒，其含有RNAi调控基因的cDNA片 段序列，cDNA片段的序列如SEQ ID NO:4所示。
需要进一步说明的，该重组表达质粒通过以下方法获得：在正向和反 向引物的5’端添加HindⅢ和XbaI的酶切位点，扩增上游片段；及在正 向和反向引物的5’端添加ApaI和SpeI的酶切位点，扩增下游片段；然后 使用HindⅢ、XbaI酶分别对质粒和上游扩增片段进行酶切，酶切后进行连 接；使用ApaI、SpeI酶对下游扩增片段和连接产物进行酶切，酶切后进 行连接。
需要进一步说明的，所述质粒为pDH-RH质粒。
本发明还提供了一种重组表达菌株，该重组表达菌株以农杆菌为宿主 细胞，将含有RNAi调控基因的cDNA片段序列的重组表达质粒转入到宿 主细胞得到。
本发明还提供所述的松材线虫RNAi调控基因在制备松材线虫防治菌 剂中的应用。需要进一步说明的，所述防治菌剂为转化有RNAi调控基因 的真菌。
需要进一步说明的，所述真菌为尖孢镰刀菌。
本发明通过农杆菌介导转化丝状真菌尖孢镰刀菌(Fusarium oxysporum)中，用阳性转化子饲喂松材线虫，获得明显的RNAi表型， 松材线虫Bx-etr-1基因表达明显下调，线虫出现胚胎发育不正常，幼虫死 亡率高，运动轨迹改变，运动速度变慢，种群数量明显下降的现象。因此， 可将本发明得到的松材线虫RNAi调控基因应用于松材线虫的防控中。
本发明的有益效果为：
本发明筛选到一个高效的RNAi调控基因，将含有该基因的阳性转化 子饲喂松材线虫，获得明显的RNAi表型，松材线虫Bx-etr-1基因表达明 显下调，松材线虫出现胚胎发育不正常，幼虫死亡率高，运动轨迹改变， 运动速度变慢，种群数量明显下降的现象。
附图说明
图1为秀丽隐杆线虫ETR蛋白的结构域；
图2为本发明靶标基因RNAi片段的PCR产物电泳图谱；
图3为本发明重组表达质粒的上下游片段的PCR产物电泳图谱；
图4为本发明尖孢镰刀菌阳性转化株生长在含抗生素的PDA平板示意 图；
图5为本发明检测Bx-etr-1基因插入阳性转化株基因组的Southern blotting图；
图6为本发明检测Bx-etr-1基因在阳性转化株中的表达的RT-PCR图；
图7为本发明检测取食阳性转化株后松材线虫的etr-1基因表达水平的 RT-qPCR图；
图8为本发明RNAi Bx-etr-1基因对松材线虫运动速率的影响；
图9为本发明RNAi Bx-etr-1基因对松材线虫幼虫的存活率的影响；
图10为本发明RNAi Bx-etr-1基因改变松材线虫的运动轨迹；其中， (A-B)：野生型松材线虫运动轨迹；(C-D)：dsGFP松材线虫运动轨迹； (E-M)：干扰后松材线虫运动轨迹；
图11为本发明RNAi Bx-etr-1基因影响松材线虫的胚胎发育；其中， (A-C)：饲喂野生型尖孢镰刀菌的松材线虫卵；(D-F)：饲喂dsGFP转 化菌的松材线虫卵；(G-I)：饲喂转化菌etr-1-1的线虫卵；(J-L)：饲喂 转化菌etr-1-2的线虫卵；(M-O)：饲喂转化菌etr-1-3的线虫卵；
图12为本发明RNAi Bx-etr-1基因对松材线虫种群数量的影响；其中， (A-C)：饲喂野生型尖孢镰刀菌的松材线虫；(D-F)：饲喂dsGFP转化 菌的松材线虫；(G-I)：饲喂转化菌etr-1-1的线虫；(J-L)：饲喂转化菌的线虫；(M-O)：饲喂转化菌etr-1-3的线虫。
具体实施方式
下面结合附图和实施例，对本发明进行具体描述。
实施例1
在NCBI网站上，通过BLAST分析，先找到与秀丽隐杆线虫ETR-1蛋 白有最高同源的蛋白，该蛋白的氨基酸序列如SEQ ID NO:3所示，秀丽隐 杆线虫ETR蛋白的结构域如图1所示。然后BLAST分析，在松材线虫基因 组找到编码SEQ ID NO:3所示氨基酸序列的基因全长序列(如SEQ ID NO:1所示)及cDNA全长序列(如SEQ ID NO:2所示)。通过PCR和 RT-PCR对序列测定验证，获得Bx-etr-1基因全长(2748bp)和cDNA全长(1539bp)，其编码蛋白大小为512aa。
采用Trizol或RNA提取试剂盒提取松材线虫总RNA。按照PrimeScript TM II 1ststrand cDNA Synthesis Kit(TaKaRa，Japan)说明进行cDNA第一 链的合成。设计etr-1基因干扰片段的正反向引物etr-1-F/R，如SEQ ID NO:5、 SEQ ID NO:6所示(见表1)，以cDNA为模板，PCR扩增靶标基因，如 图1所示，获得900bp的cDNA片段序列(SEQ ID NO:4所示)。用Axygen 凝胶回收试剂盒进行切胶回收后，测序验证。
表1
实施例2
含有cDNA片段序列的重组表达载体的构建方法包括以下步骤：
(1)在正向引物etr-1-F和反向引物etr-1-R的5’端分别添加HindⅢ和 XbaI(HindⅢ-etr-1-F/XbaI-etr-1-R)，扩增上游片段；及在正向引物etr-1-F 和反向引物etr-1-R的5’端分别添加ApaI和SpeI的酶切位点序列(ApaI -etr-1-F/SpeI-etr-1-R)(见表1)，扩增下游片段，RT-PCR扩增的反应体系 如表2所示。
表2
反应条件为
(2)使用HindⅢ、XbaI酶分别对质粒和上游扩增片段进行酶切，酶切 后进行连接；然后使用ApaI、SpeI酶对下游扩增片段和连接产物进行 酶切，酶切后进行连接。将pDH-RH质粒和扩增片段分别都用HindⅢ和 XbaI，以及ApaI和SpeI进行双酶切后，采用Rapid DNAligation Kit将酶 切后的质粒和扩增片段进行连接反应。先将一个片段插入载体中，转化到 大肠杆菌HST02感受态细胞后，在含有100mg/ml卡那霉素的LB平板上 进行阳性克隆的筛选和PCR检测。鉴定后的阳性克隆质粒用于另一个片 段的插入，同样的方法筛选阳性克隆。提取最终获得的阳性单克隆质粒， 进行正向和反向插入片段的PCR检测，如图2所示，得到具有双向etr-1 插入片段的载体为正确的能表达Bx-etr-1基因dsRNA的载体。上游酶切反应体系如表3所示，下游酶切反应体系如表4所示，分别在37℃，酶切 12-16h。连接反应体系如表5所示，22℃，连接20min。
表3
表4
表5
实施例3
重组表达菌株的构建方法包括以下步骤：将实施例2获得的正确表达 Bx-etr-1基因dsRNA的质粒体转化到农杆菌中，在含有卡那霉素(100mg/L) 和利福平(50mg/L)的LB平板上筛选阳性克隆。提取阳性菌株的质粒，用 上下游引物PCR扩增进行分子鉴定，得到正反向插入片段的阳性菌株。
实施例4
将实施例3得到的含有正反向插入片段的阳性菌株用于转染受体真菌分 子孢子。将受体真菌尖孢镰刀菌分生孢子和上述阳性农杆菌株分别用IM培 养液悬浮后，等量混合，并加入200μmol/L乙酰丁香酮(AS)，避光培养(25℃， 110r/min)48h后涂布于铺有灭菌滤纸的IM平板上培养。待真菌孢子萌发后， 将滤纸反铺到含有100mg/L的潮霉素B和100mg/L的头孢霉素的MM平板 上培养7-10天。待尖孢镰刀菌长出后，挑至含有上述抗生素的PDA平板上 培养，如图4所示，连续转接5-7次后，进行单孢分离和培养。
收集尖孢镰刀菌阳性菌株的菌丝，提取基因组DNA，通过PCR扩增靶 标基因的上下游片段进行快速分子鉴定。同时，将靶标基因片段合成探针， 通过Southern杂交,确定T-DNA在真菌基因组中的随机插入及拷贝数，如 图5所示，T-DNA在真菌基因组中为单拷贝数插入。提取尖孢镰刀菌阳性菌 株RNA，合成cDNA第一链，以此为模板，用etr-1-F/R引物进行RT-PCR 扩增，检测etr-1基因在尖孢镰刀菌基因组中的表达，如图6所示。
实施例5
经上述分子鉴定的尖孢镰刀菌转化株培养在PDA平板上，4-5天后接种 松材线虫，培养20天后，进行RNAi表型观察。提取松材线虫RNA，合成 cDNA第一链，以qRT-etr-1-F/R为引物(见表1)，RT-qPCR检测，发现松 材线虫靶标基因的敲低65％以上，如图7所示。松材线虫卵和幼虫出现明显 的异常，表现在胚胎死亡(如图11所示)，幼虫运动速率降低(如图8所示)，运动轨迹改变(如图10所示)，死亡率明显达50％左右(如图9所示)，种 群数量减少(如图12所示)。与对照相比，差异显著。
上述实施例中无特殊说明的操作手法均为现有技术，故不在此过多解 释。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本 领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和 原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护 范围之内。
序列表
<110> 北京师范大学
<120> 一种松材线虫RNAi调控基因及其应用
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<170> SIPOSequenceListing 1.0
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atggggctcc aatgtgaaga ggccataaac cccaatcctg atgtggacac ggccaggccc 60
tccagcaccg atagtaatgg gtttccggtt aaagattcag atgcaattaa gctgttcgtc 120
ggccaagtaa gttcagtttt atgcatcatc tcttccaaaa aacagtctgc ggtatcggaa 180
atgggcagag agactgcaac ccattaattt ggattattta ttttgcataa ttaattcggc 240
cttaatattt ctttgtaaat ttccaaaaat gcccgaaggt ttaagacttt ctggctttcg 300
tatgtctata cgaaagcagt aagcttcgta tagatattca taaagaatac ttttactatt 360
accaagtaat caacgggtat ttagggaggg gagatttgcg tcatggggac cacagtaaaa 420
taagtactta tacgcctcga aaacataaaa aacgtcaata tttggctcga gaagtgcatt 480
tgaatcgcca ccaataattg ttttcttttt cagataccac ggaatctgga ggagaaagac 540
ctccgacata tgttcgagac cttcgggaag atctacgagt tcacaatcct caaggataaa 600
tacacgggca tgcacaaagg ctgcgccttc ctgacctact gtcatcgaga ccacgctctc 660
cgatgtcagg ccgccctcca cgatcagaaa acccttccag gagtaagtta tttgggccta 720
aagtggccga ccaaacttat ttagggaggg accttgagaa gtgcatcaaa tttgggaaga 780
aaacgcccta aggcactgaa tttggtgcaa tttgggtaag aaaaagggga aacgaggccg 840
gggccggtgc gaggccggag acaaacaatt ccgaagaaag tgtggccgca aattcaaggt 900
tcactacctc cgtttggaca ctttggaggg aaagtttact ttattctact gactttcaga 960
tgaaccgtgc catgcaagtg aagccggcgg actcggattc tcgccccaat tcccccaaac 1020
cgagtgatga tcgaaagttg ttcattggaa tgttatccaa gcaacaagga gaagaggaag 1080
tcaaggcact cctgtcaccg ttcgggaaga tcgaagaagt tacagtacta cgagctgctg 1140
atggagtttc caaggtaagg ctcggaggca aagcttggga cattcctttg ttctttgcaa 1200
taaatccaag tttattttat cttcttcttc tccagggatg tgcctttgcc aaattcacga 1260
atgcgggcga tgcccaaagg gctatttccg ctcttcatgg aagccagacg atgagaggcg 1320
cctcgagcag tcttgtggtg aagttggcgg acaccgacaa ggaaagacag ctcaggagaa 1380
tgcagcagat ggcttcgcag attgggattc tgaaccccct tctggccgcc caatcgggaa 1440
tttacggtgc agctacggcc agtcctctaa accagttaaa tgtaagcaaa ccaaagagtc 1500
ttaattcagt ttgaaggtgt tgcaacaaca acaagcagct caattggtgg ccgcggcttc 1560
catacaaagc ccggtaacgg cttcgtatct tccattactg cagagtcaag ccttgagtac 1620
tccaaataac attctcccaa cccagagttc aacgaatcaa gccgccgttg cagcggccgt 1680
tgctgcagct caggcacaag ttcaagcagc ggcgcagatt caacacctcg gtcttcagag 1740
ccagattccg gcttcgaatc cacaactaca aagtcagtta tcagctttgg cagcccatca 1800
agggaccacc tcgactgcta attccaacgg ggagttaaca acgactcagg cctacggggt 1860
ccaagctggc gcctataacc agttgattgc ctatgaccaa tcgcagaatt gtaaggatta 1920
tctgaccctg atagaacgct ttcagcttca gccatgtaca accaaaccat ccagcaatta 1980
caacaacaac aaatggccca attggcagcc gttgccgccg cctcaggcgc tgggattctc 2040
cccggtttca cgcataaaga aggtaaggga aagaggtccg atactaatgt tgttttagtc 2100
cccggaccgg aagggtgtaa cctcttcatc taccatcttc ctcaagagtt tggagatgct 2160
gaattgacac aaatgttttt gcctttcggc accgtgttga gtgccaaagt cttcattgat 2220
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cttttttgac gccgctctca gcggattcag agtactgtaa ctatagttaa ttgattggtt 2400
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ggatgacaac tttgagtatt gtaatgccaa acacggagtt ctcctttttt gagagtgagc 2520
acagactaga ttatattatc cttgagagtt gttcctcgtt catttctgat gcccattaag 2580
tccgaggcta gtacttgttt ctcttttttt gtacagtgtt ttctaggatt tgtatcgtac 2640
gataacccca actcggcgat ggcggcaatc caggcgatga atgggttcca gatcgggatg 2700
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<210> 2
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ggccaaatac cacggaatct ggaggagaaa gacctccgac atatgttcga gaccttcggg 180
aagatctacg agttcacaat cctcaaggat aaatacacgg gcatgcacaa aggctgcgcc 240
ttcctgacct actgtcatcg agaccacgct ctccgatgtc aggccgccct ccacgatcag 300
aaaacccttc caggaatgaa ccgtgccatg caagtgaagc cggcggactc ggattctcgc 360
cccaattccc ccaaaccgag tgatgatcga aagttgttca ttggaatgtt atccaagcaa 420
caaggagaag aggaagtcaa ggcactcctg tcaccgttcg ggaagatcga agaagttaca 480
gtactacgag ctgctgatgg agtttccaag ggatgtgcct ttgccaaatt cacgaatgcg 540
ggcgatgccc aaagggctat ttccgctctt catggaagcc agacgatgag aggcgcctcg 600
agcagtcttg tggtgaagtt ggcggacacc gacaaggaaa gacagctcag gagaatgcag 660
cagatggctt cgcagattgg gattctgaac ccccttctgg ccgcccaatc gggaatttac 720
ggtgcagcta cggccagtcc tctaaaccag ttaaatgtgt tgcaacaaca acaagcagct 780
caattggtgg ccgcggcttc catacaaagc ccggtaacgg cttcgtatct tccattactg 840
cagagtcaag ccttgagtac tccaaataac attctcccaa cccagagttc aacgaatcaa 900
gccgccgttg cagcggccgt tgctgcagct caggcacaag ttcaagcagc ggcgcagatt 960
caacacctcg gtcttcagag ccagattccg gcttcgaatc cacaactaca aagtcagtta 1020
tcagctttgg cagcccatca agggaccacc tcgactgcta attccaacgg ggagttaaca 1080
acgactcagg cctacggggt ccaagctggc gcctataacc agttgattgc ctatgaccaa 1140
tcgcagaatt cttcagccat gtacaaccaa accatccagc aattacaaca acaacaaatg 1200
gcccaattgg cagccgttgc cgccgcctca ggcgctggga ttctccccgg tttcacgcat 1260
aaagaagtcc ccggaccgga agggtgtaac ctcttcatct accatcttcc tcaagagttt 1320
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ttcattgatc gggccacaaa tcagagcaaa tgtttcggat ttgtatcgta cgataacccc 1440
aactcggcga tggcggcaat ccaggcgatg aatgggttcc agatcgggat gaaacgcctc 1500
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<210> 3
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1 5 10 15
Thr Ala Arg Pro Ser Ser Thr Asp Ser Asn Gly Phe Pro Val Lys Asp
20 25 30
Ser Asp Ala Ile Lys Leu Phe Val Gly Gln Ile Pro Arg Asn Leu Glu
35 40 45
Glu Lys Asp Leu Arg His Met Phe Glu Thr Phe Gly Lys Ile Tyr Glu
50 55 60
Phe Thr Ile Leu Lys Asp Lys Tyr Thr Gly Met His Lys Gly Cys Ala
65 70 75 80
Phe Leu Thr Tyr Cys His Arg Asp His Ala Leu Arg Cys Gln Ala Ala
85 90 95
Leu His Asp Gln Lys Thr Leu Pro Gly Met Asn Arg Ala Met Gln Val
100 105 110
Lys Pro Ala Asp Ser Asp Ser Arg Pro Asn Ser Pro Lys Pro Ser Asp
115 120 125
Asp Arg Lys Leu Phe Ile Gly Met Leu Ser Lys Gln Gln Gly Glu Glu
130 135 140
Glu Val Lys Ala Leu Leu Ser Pro Phe Gly Lys Ile Glu Glu Val Thr
145 150 155 160
Val Leu Arg Ala Ala Asp Gly Val Ser Lys Gly Cys Ala Phe Ala Lys
165 170 175
Phe Thr Asn Ala Gly Asp Ala Gln Arg Ala Ile Ser Ala Leu His Gly
180 185 190
Ser Gln Thr Met Arg Gly Ala Ser Ser Ser Leu Val Val Lys Leu Ala
195 200 205
Asp Thr Asp Lys Glu Arg Gln Leu Arg Arg Met Gln Gln Met Ala Ser
210 215 220
Gln Ile Gly Ile Leu Asn Pro Leu Leu Ala Ala Gln Ser Gly Ile Tyr
225 230 235 240
Gly Ala Ala Thr Ala Ser Pro Leu Asn Gln Leu Asn Val Leu Gln Gln
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Gln Gln Ala Ala Gln Leu Val Ala Ala Ala Ser Ile Gln Ser Pro Val
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Thr Ala Ser Tyr Leu Pro Leu Leu Gln Ser Gln Ala Leu Ser Thr Pro
275 280 285
Asn Asn Ile Leu Pro Thr Gln Ser Ser Thr Asn Gln Ala Ala Val Ala
290 295 300
Ala Ala Val Ala Ala Ala Gln Ala Gln Val Gln Ala Ala Ala Gln Ile
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Gln His Leu Gly Leu Gln Ser Gln Ile Pro Ala Ser Asn Pro Gln Leu
325 330 335
Gln Ser Gln Leu Ser Ala Leu Ala Ala His Gln Gly Thr Thr Ser Thr
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Ala Asn Ser Asn Gly Glu Leu Thr Thr Thr Gln Ala Tyr Gly Val Gln
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Ala Gly Ala Tyr Asn Gln Leu Ile Ala Tyr Asp Gln Ser Gln Asn Ser
370 375 380
Ser Ala Met Tyr Asn Gln Thr Ile Gln Gln Leu Gln Gln Gln Gln Met
385 390 395 400
Ala Gln Leu Ala Ala Val Ala Ala Ala Ser Gly Ala Gly Ile Leu Pro
405 410 415
Gly Phe Thr His Lys Glu Val Pro Gly Pro Glu Gly Cys Asn Leu Phe
420 425 430
Ile Tyr His Leu Pro Gln Glu Phe Gly Asp Ala Glu Leu Thr Gln Met
435 440 445
Phe Leu Pro Phe Gly Thr Val Leu Ser Ala Lys Val Phe Ile Asp Arg
450 455 460
Ala Thr Asn Gln Ser Lys Cys Phe Gly Phe Val Ser Tyr Asp Asn Pro
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Asn Ser Ala Met Ala Ala Ile Gln Ala Met Asn Gly Phe Gln Ile Gly
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<210> 4
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<212> DNA
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gggctatttc cgctcttcat ggaagccaga cgatgagagg cgcctcgagc agtcttgtgg 180
tgaagttggc ggacaccgac aaggaaagac agctcaggag aatgcagcag atggcttcgc 240
agattgggat tctgaacccc cttctggccg cccaatcggg aatttacggt gcagctacgg 300
ccagtcctct aaaccagtta aatgtgttgc aacaacaaca agcagctcaa ttggtggccg 360
cggcttccat acaaagcccg gtaacggctt cgtatcttcc attactgcag agtcaagcct 420
tgagtactcc aaataacatt ctcccaaccc agagttcaac gaatcaagcc gccgttgcag 480
cggccgttgc tgcagctcag gcacaagttc aagcagcggc gcagattcaa cacctcggtc 540
ttcagagcca gattccggct tcgaatccac aactacaaag tcagttatca gctttggcag 600
cccatcaagg gaccacctcg actgctaatt ccaacgggga gttaacaacg actcaggcct 660
acggggtcca agctggcgcc tataaccagt tgattgccta tgaccaatcg cagaattctt 720
cagccatgta caaccaaacc atccagcaat tacaacaaca acaaatggcc caattggcag 780
ccgttgccgc cgcctcaggc gctgggattc tccccggttt cacgcataaa gaagtccccg 840
gaccggaagg gtgtaacctc ttcatctacc atcttcctca agagtttgga gatgctgaat 900
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Claims (10)
1.一种松材线虫RNAi调控基因，其特征在于，该RNAi调控基因的基因全长序列如SEQID NO:1所示。
2.根据权利要求1所述的松材线虫RNAi调控基因，其特征在于，该RNAi调控基因的cDNA全长序列如SEQ ID NO:2所示。
3.根据权利要求1所述的松材线虫RNAi调控基因，其特征在于，该RNAi调控基因的编码氨基酸序列如SEQ ID NO:3所示。
4.一种重组表达质粒，其特征在于，其含有RNAi调控基因的cDNA片段序列，cDNA片段的序列如SEQ ID NO:4所示。
5.根据权利要求4所述的重组表达质粒，其特征在于，该重组表达质粒通过以下方法获得：在正向和反向引物的5’端添加HindⅢ和XbaI的酶切位点，扩增上游片段；及在正向和反向引物的5’端添加Apa|和SpeI的酶切位点，扩增下游片段；然后使用HindⅢ、Xba|酶分别对质粒和上游扩增片段进行酶切，酶切后进行连接；使用ApaI、SpeI酶对下游扩增片段和连接产物进行酶切，酶切后进行连接。
6.根据权利要求5所述的重组表达质粒，其特征在于，所述质粒为pDH-RH质粒。
7.一种重组表达菌株，其特征在于，该重组表达菌株以农杆菌为宿主细胞，将含有RNAi调控基因的cDNA片段序列的重组表达质粒转入到宿主细胞得到。
8.权利要求1所述的松材线虫RNAi调控基因在制备松材线虫防治菌剂中的应用。
9.根据权利要求8所述的应用，其特征在于，所述防治菌剂为转化有RNAi调控基因的真菌。
10.根据权利要求9所述的应用，其特征在于，所述真菌为尖孢镰刀菌。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257407A (zh) * | 2019-07-08 | 2019-09-20 | 东北林业大学 | 一种海藻糖酶基因Bx-tre1及其应用 |
| CN110283827A (zh) * | 2019-07-17 | 2019-09-27 | 山东农业大学 | 松材线虫mog-2基因及其在发育干扰中应用 |
| CN110777149A (zh) * | 2019-10-15 | 2020-02-11 | 山东农业大学 | 松材线虫tra-1基因及其在发育干扰中的应用 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040213771A1 (en) * | 2001-03-20 | 2004-10-28 | Sluder Ann E | Targets and screens for agents useful in controlling parasitic nematodes |
| CN102286509A (zh) * | 2011-06-27 | 2011-12-21 | 中国农业科学院蔬菜花卉研究所 | 干扰Bx-cpl-1基因表达的dsRNA载体及其在松材线虫防治和研究中的应用 |
| CN102250942B (zh) * | 2011-06-27 | 2013-08-14 | 中国农业科学院蔬菜花卉研究所 | 干扰Bx-cpl-2基因表达的dsRNA载体及其在松材线虫防治和研究中的应用 |
| CN106755055A (zh) * | 2017-03-08 | 2017-05-31 | 中国农业大学 | 用于抑制松材线虫基因表达的成套试剂与方法 |
| CN107557365A (zh) * | 2017-08-31 | 2018-01-09 | 长江师范学院 | 一种双元靶标dsRNA及合成方法和应用 |
-
2018
- 2018-12-28 CN CN201811624288.4A patent/CN109402171B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040213771A1 (en) * | 2001-03-20 | 2004-10-28 | Sluder Ann E | Targets and screens for agents useful in controlling parasitic nematodes |
| CN102286509A (zh) * | 2011-06-27 | 2011-12-21 | 中国农业科学院蔬菜花卉研究所 | 干扰Bx-cpl-1基因表达的dsRNA载体及其在松材线虫防治和研究中的应用 |
| CN102250942B (zh) * | 2011-06-27 | 2013-08-14 | 中国农业科学院蔬菜花卉研究所 | 干扰Bx-cpl-2基因表达的dsRNA载体及其在松材线虫防治和研究中的应用 |
| CN106755055A (zh) * | 2017-03-08 | 2017-05-31 | 中国农业大学 | 用于抑制松材线虫基因表达的成套试剂与方法 |
| CN107557365A (zh) * | 2017-08-31 | 2018-01-09 | 长江师范学院 | 一种双元靶标dsRNA及合成方法和应用 |
Non-Patent Citations (2)
| Title |
|---|
| 戴素明等: "松材线虫Hsp70基因的克隆与原核表达", 《植物病理学报》 * |
| 王殿东等: "松材线虫mapk基因克隆及RNAi效应分析", 《植物病理学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257407A (zh) * | 2019-07-08 | 2019-09-20 | 东北林业大学 | 一种海藻糖酶基因Bx-tre1及其应用 |
| CN110257407B (zh) * | 2019-07-08 | 2023-04-28 | 东北林业大学 | 一种海藻糖酶基因Bx-tre1及其应用 |
| CN110283827A (zh) * | 2019-07-17 | 2019-09-27 | 山东农业大学 | 松材线虫mog-2基因及其在发育干扰中应用 |
| CN110283827B (zh) * | 2019-07-17 | 2021-06-08 | 山东农业大学 | 松材线虫mog-2基因及其在发育干扰中应用 |
| CN110777149A (zh) * | 2019-10-15 | 2020-02-11 | 山东农业大学 | 松材线虫tra-1基因及其在发育干扰中的应用 |
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