CN112538490A - 一种生防腐霉菌中诱导坏死和活性氧积累的nlp类基因及其表达载体和应用 - Google Patents

Abstract

本发明属于基因工程技术领域，公开了一种生防腐霉菌诱导坏死和活性氧积累的NLP类基因及其表达载体和应用。本发明首次从寡雄腐霉菌和缠器腐霉菌两种生防腐霉菌中鉴定出11个NLP类基因，该11个NLP类基因PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8基因的核苷酸序列依次分别如SEQ ID NO.1—11所示。该基因能够诱导植物产生显著的坏死，促进活性氧积累。该NLP基因可以作为开发植物免疫诱抗剂的潜在候选者，为防控植物病害理论支撑。

Description

一种生防腐霉菌中诱导坏死和活性氧积累的NLP类基因及其 表达载体和应用

技术领域

本发明属于基因工程技术领域，具体涉及一种生防腐霉菌诱导坏死和活性氧积累的NLP类基因及其表达载体和应用。

背景技术

寡雄腐霉菌(Pythium oligandrum)和缠器腐霉菌(Pythium periplocum)广泛存在于植物根系生态系统，在与植物接触过程中通过分泌大量效应子诱导植物对病原菌产生广泛、长期的抗性，同时对很多致病菌有很强的重寄生和拮抗作用。

坏死与乙烯诱导肽类蛋白(NLPs)广泛存在于各种植物致病微生物中。在其遗传进化上不同于免疫相关的程序性死亡，并且在双子叶植物中能够诱导坏死，而在单子叶植物中不能诱导坏死。因此，研究人员提出NLP类效应蛋白具有双重功能，既能触发免疫反应，又能行使毒性功能。我们首次鉴定寡雄腐霉菌和缠器腐霉菌两种生防腐霉菌中存在NLP类效应蛋白，并且其具有诱导植物产生坏死和活性氧积累的强大功能。活性氧积累是一个触发植物免疫反应的关键信号，也可以作为植物防御反应触发的信号标识。因此本发明中鉴定的NLP类蛋白具有开发成为植物免疫诱抗剂的潜质。

发明内容

本发明的目的在于提供一种生防腐霉菌中诱导坏死和活性氧积累的NLP类基因及其编码的蛋白和应用。本发明鉴定到的11个NLP类基因可以诱导坏死和活性氧积累。

本发明的目的可以通过以下技术方案实现：

本发明首次从寡雄腐霉菌和缠器腐霉菌两种生防腐霉菌中鉴定出11个NLP类基因，并且研究发现其具有诱导植物产生坏死和活性氧积累的强大功能。该11个NLP类基因包括分别来自寡雄腐霉(Pythium oligandrum)的8个NLP类基因PyolNLP1/PyolNLP2/PyolNLP3/PyolNLP4/PyolNLP5/PyolNLP6/PyolNLP7/PyolNLP9和缠器腐霉(Pythiumperiplocum)的3个NLP类基因PypeNLP4/PypeNLP5/PypeNLP8，上述NLP类基因PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8的核苷酸序列依次分别如SEQ ID NO.1—11所示。

上述NLP类基因PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8编码的蛋白质，其氨基酸序列依次分别如SEQ ID NO.12—22所示。本发明将从寡雄腐霉和缠器腐霉中鉴定到11个NLP类基因分别插入表达载体PBIN-PLUS，并将所述载体导入植物中，均能够产生显著的坏死，促进活性氧积累。蛋白表达进行诱导植物抗性实验证明，其中PyolNLP5和PyolNLP7诱导植物抗性表型最好，在烟草、番茄、辣椒上能够发挥对多种病原菌的抗病表型，这两个蛋白具有更高的开发成生防产品的潜质。

含有上述NLP类基因的表达盒、重组表达载体、转基因细胞系或转基因重组菌。

上述重组表达载体的出发载体为表达载体PBIN-PLUS。含有上述NLP类基因的重组表达载体PBIN-PLUS：PyolNLP1、PBIN-PLUS：PyolNLP2、PBIN-PLUS：PyolNLP3、PBIN-PLUS：PyolNLP4、PBIN-PLUS：PyolNLP5、PBIN-PLUS：PyolNLP6、PBIN-PLUS：PyolNLP7、PBIN-PLUS：PyolNLP9、PBIN-PLUS：PypeNLP4、PBIN-PLUS：PypeNLP5和PBIN-PLUS：PypeNLP8。该重组表达载体为将上述的NLP类基因插入PBIN-PLUS载体的SmaI酶切位点间所得。

上述的NLP类基因、蛋白或重组表达载体在诱导植物产生坏死和活性氧积累中的应用。

上述的NLP类基因、蛋白或重组表达载体在开发植物免疫诱抗剂中的应用。

本发明的有益效果：

本发明分别从寡雄腐霉和缠器腐霉中鉴定到11个NLP类基因，将其插入表达载体PBIN-PLUS。将所述载体导入植物中，能够产生显著的坏死，促进活性氧积累。所述NLP基因可以作为开发植物免疫诱抗剂的潜在候选者，为防控植物病害理论支撑。

附图说明

图1为NLP类基因诱导植物坏死和活性氧积累显著效果的表型图。

其中，顺序如图标识，GFP处理为阴性对照，INF1处理为阳性对照。

图2为活性氧迸发强度定量统计图。

图3为PyolNLP5和PyolNLP7蛋白在本式烟中诱导对辣椒疫霉和寄生疫霉的抗性。

图4为PyolNLP5和PyolNLP7蛋白在辣椒中诱导对辣椒疫霉的抗性。

图5为PyolNLP5和PyolNLP7蛋白在番茄中诱导对辣椒疫霉的抗性。

具体实施方式

以下结合具体实施例对本发明做出详细的描述。根据以下的描述和这些实施例，本领域技术人员可以确定本发明的基本特征，并且在不偏离本发明精神和范围的情况下，可以对本发明做出各种改变和修改，以使其适用各种用途和条件。

实施例1 11个NLP类基因的扩增和测序

1.实验菌株

供试菌株寡雄腐霉(Pythium oligandrum)¹和缠器腐霉(Pythium periplocum)²由南京农业大学植病系植物与疫霉互作实验室保存菌株保存于10％V8固体培养基，保存温度为10℃。

2.供试本氏烟草幼苗的准备

本氏烟草(Nicotiana benthamiana)播种于装有蛭石(2-4mm)的塑料花盆(d＝10cm)里，放到14h光照(强光照射)/10h黑暗的温室中生长。使其生长7天，生长出两片真叶后，将其移栽至蛭石黑土体积比例为5：1的培养土里生长30天，取相同3，4，5叶位的叶片用于表达实验基因，然后持续观察7天叶片表型是否有明显坏死。

3.提取寡雄腐霉(Pythium oligandrum)和缠器腐霉(Pythium periplocum)RNA，并获得目的基因的cDNA

收集寡雄腐霉和缠器腐霉菌丝，然后液氮速冻后放入研钵冷却研磨。参照天根公司总RNA抽提纯化试剂盒推荐方法进行寡雄腐霉和缠器腐霉总RNA提取和纯化。参照诺唯赞公司RNA反转录试剂盒提供的方法，以Oligo(dT)为引物反转录合成cDNA。以上述cDNA为模板，用相应上下游引物(引物序列见表1)进行PCR扩增并获得11个NLP类基因。

表1 引物序列

PCR反应体系为：2.5μL 10×PCR反应缓冲液；1.5μL 1.5mM MgCl₂；0.5μL 2.5mMdNTPs；0.25μL Taq DNA聚合酶(5.0U/μL)；0.5μL引物；0.5μL模板；无菌水补足至25μL。

反应程序为：94℃5min预变性；94℃变性15s，58℃退火15s，72℃延伸1:30min，35个循环；72℃延伸10min；4℃保存。

上述PCR扩增产物经琼脂糖凝胶电泳、回收，经测序确认无误后，获得的序列为所述11个NLP类基因的核苷酸序列。该11个NLP类基因包括分别来自寡雄腐霉(Pythiumoligandrum)的8个NLP类基因PyolNLP1/PyolNLP2/PyolNLP3/PyolNLP4/PyolNLP5/PyolNLP6/PyolNLP9和缠器腐霉(Pythium periplocum)的3个NLP类基因PypeNLP4/PypeNLP5/PypeNLP8，上述NLP类基因PyolNLP1/PyolNLP2/PyolNLP3/PyolNLP4/PyolNLP5/PyolNLP6/PyolNLP7/PyolNLP9和PypeNLP4/PypeNLP5/PypeNLP8的核苷酸序列依次如SEQID NO.1—11所示。后续研究发现其具有诱导植物产生坏死和活性氧积累的强大功能。

上述NLP类基因PyolNLP1/PyolNLP2/PyolNLP3/PyolNLP4/PyolNLP5/PyolNLP6/PyolNLP7/PyolNLP9和PypeNLP4/PypeNLP5/PypeNLP8编码的蛋白质，其氨基酸序列为SEQID NO.12—22。

实施例2构建PBIN-PLUS:PyolNLP1/PyolNLP2/PyolNLP3/PyolNLP4/PyolNLP5/PyolNLP6/PyolNLP7/PyolNLP9/PypeNLP4/PypeNLP5/PypeNLP8表达载体

将PBIN-PLUS(BIOVECTOR中国质粒载体菌株细胞株基因保藏中心)空载体质粒用SmaI进行酶切。利用诺唯赞公司的同源重组酶将实施例1获得的PCR扩增产物(11个NLP类基因PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8)分别与载体连接。

反应体系如下：2μL 10×CEⅡ反应缓冲液；2μL上述PCR回收产物；2μL上述酶切后的空载体；1μL同源重组酶；无菌水补足至10μL。

反应程序为：37℃条件下反应30min。将连接产物转化大肠杆菌DH5α，在Kan抗性培养基上筛选转化子，挑取阳性克隆提取质粒进行菌落PCR鉴定。将鉴定到的阳性克隆测序，即正确获得了PBIN-PLUS：PyolNLP1、PBIN-PLUS：PyolNLP2、PBIN-PLUS：PyolNLP3、PBIN-PLUS：PyolNLP4、PBIN-PLUS：PyolNLP5、PBIN-PLUS：PyolNLP6、PBIN-PLUS：PyolNLP7、PBIN-PLUS：PyolNLP9、PBIN-PLUS：PypeNLP4、PBIN-PLUS：PypeNLP5、PBIN-PLUS：PypeNLP8共11个表达载体。

实施例3表达载体转化农杆菌

1.获得农杆菌感受态细胞

在LB固体培养基平板上划线培养农杆菌GV3101菌株，28℃条件下倒置培养18-20h；将单菌落置于LB液体培养基(含有100mg/L Rif)，28℃条件下振荡培养18-20h；100倍体积的不含抗生素的菌液加入菌液中，28℃条件下振荡培养OD600至0.3-0.5；冰上冷却后，4℃条件下4000r/min离心5min，弃上清；加入20mmol/L的CaCl₂溶液使沉淀悬浮，4℃条件下4000r/min离心5min，弃上清；加入20mmol/L的CaCl₂溶液再次使菌体沉淀悬浮备用。

2.PBIN-PLUS：PyolNLP1、PBIN-PLUS：PyolNLP2、PBIN-PLUS：PyolNLP3、PBIN-PLUS：PyolNLP4、PBIN-PLUS：PyolNLP5、PBIN-PLUS：PyolNLP6、PBIN-PLUS：PyolNLP7、PBIN-PLUS：PyolNLP9、PBIN-PLUS：PypeNLP4、PBIN-PLUS：PypeNLP5、PBIN-PLUS：PypeNLP8表达载体转化农杆菌

利用电转法将实施例2制备的PBIN-PLUS：PyolNLP1、PBIN-PLUS：PyolNLP2、PBIN-PLUS：PyolNLP3、PBIN-PLUS：PyolNLP4、PBIN-PLUS：PyolNLP5、PBIN-PLUS：PyolNLP6、PBIN-PLUS：PyolNLP7、PBIN-PLUS：PyolNLP9、PBIN-PLUS：PypeNLP4、PBIN-PLUS：PypeNLP5、PBIN-PLUS：PypeNLP8载体分别转化农杆菌细胞；利用70％酒精冲洗电击杯三次，然后完全风干酒精。将100ng表达载体加入农杆菌感受态，冰上放置30min。将农杆菌感受态细胞转移至电击杯中，利用2.5kV的电压进行电击转化。电击完成后将农杆菌感受态细胞加入到400μL LB液体培养基中，28℃振荡培养2h。吸取20μL菌液，均匀涂抹在含有50mM卡那霉素和25mM的利福平固体LB培养基平板上，28℃倒置培养48h。挑取单菌落进行菌落PCR，获取阳性克隆。

3.在本氏烟草中分别瞬时表达PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8蛋白

将上述阳性克隆放置于LB液体培养基中，28℃震荡培养30h。将菌液收集，用8000rpm离心2min，利用10mM MgCl₂清洗三次。最后用10mM的MgCl₂稀释菌液至OD600值0.3。将菌液与含有沉默抑制子P19的菌液1：1体积比混合。用1mL注射器在本式烟叶片上分别注射菌液，每个最少5个重复，实验重复三次。

4.活性氧(ROS)积累检测³

注射所有处理菌液36h后，用1mg/ml DAB溶液于黑暗中染色8h，然后用95％乙醇脱色处理，最后用相机拍照记录表型，检测结果见图1和图2。

5.蛋白表达进行诱导植物抗性实验

蛋白表达进行诱导植物抗性实验证明，其中PyolNLP5和PyolNLP7诱导植物抗性表型最好，在烟草、番茄、辣椒上能够发挥对多种病原菌的抗病表型，这两个蛋白具有更高的开发成生防产品的潜质，实验结果见图3～5。图3结果显示PyolNLP5和PyolNLP7蛋白在本式烟中能够高效诱导对辣椒疫霉和寄生疫霉的抗性；图4结果显示PyolNLP5和PyolNLP7蛋白在辣椒中能够高效诱导对辣椒疫霉的抗性；图5结果显示PyolNLP5和PyolNLP7蛋白在番茄中能够高效诱导对辣椒疫霉的抗性。

参考文献

1.Benhamou,N.et al.Pythium oligandrum:an example of opportunisticsuccess.Microbiology 158,2679-2694,doi:10.1099/mic.0.061457-0(2012).

2.Paul,B.&Masih,I.ITS1 region of the nuclear ribosomal DNA of themycoparasite Pythium periplocum,its taxonomy,and its comparison with relatedspecies.FEMS Microbiology Letters(2000).

3.Jambunathan,N.Determination and detection of reactive oxygenspecies(ROS),lipid peroxidation,and electrolyte leakage in plants.Methods MolBiol 639,292-298,doi:10.1007/978-1-60761-702-0_18(2010).

PyolNLP1基因(810bp)

ATGGCAGTCCGTTGCTTGGTCTCTACGATCGTGCTTGCTATTGTGCCACTCGCCAGTGGGATTGGTTTCGAGGTAATCACAGGTGACGTCTTCCCGAGTCTCTCGCGCTCATGGCCCAAGCACGTGGACATCTCGTCGATTACGCCACTCTTTGACATGGATGACAATGGCTGCTACCCAGCGGCCGTCATTAGTCGCTCTGGTGAGCCGAACCGAGGCCTGGAGACCGACGGCGGCAACTATGATGTCTGTAAACGCGACGATTTCCTCGAGTACTCGAATACGTATCACCGCTACGCGTGTCGTGTCGCCGATAACATCAAGTACTGCAGTCATTTCTTCACGCTCTACTTCCAAAAAGACGAAGCAACTCTCATCGGTGGCCATCGTCACGACCTCGAGCAAGTCGTCATCTGGACCATGGACGGCAACGTCACTCACGGGAGCTTCAGCGCCCATGGTGGTATGACCACCTGGTCAGCGAGCAGCATCCCACACGAAGGCTCGCATCTCAAATTCGTGTATCACAAAGACAACATCATGACGCACGCCATGCGCTTTGCAAGGACCAAAGACGCGTTGGACCCAGAGAATCCGTACAAGAAGTTTGTGCTTCCACCAGTCGTGAGCTGGTACACGATGAAGGGCGATGGAGATCTGACCAACAGTGCGTTGCGTCAGCTTCTCAACACACTTCCTTACGGCAAAGCGACTATGCCCATCAAGGACTCCGTATTTCTGAGGAACCTCAATATGAGACGACCCCAAGGATACCCCGAGTTTGACGATGAAGACATCAAGAGCTCGCAA

PyolNLP1蛋白(270)

MAVRCLVSTIVLAIVPLASGIGFEVITGDVFPSLSRSWPKHVDISSITPLFDMDDNGCYPAAVISRSGEPNRGLETDGGNYDVCKRDDFLEYSNTYHRYACRVADNIKYCSHFFTLYFQKDEATLIGGHRHDLEQVVIWTMDGNVTHGSFSAHGGMTTWSASSIPHEGSHLKFVYHKDNIMTHAMRFARTKDALDPENPYKKFVLPPVVSWYTMKGDGDLTNSALRQLLNTLPYGKATMPIKDSVFLRNLNMRRPQGYPEFDDEDIKSSQ

PyolNLP2基因(783bp)

ATGAAGCTTTTGACGATCCTCTCCGTCGTATGCGCTGCTGTCGCTGGCTCTGTGCGCGCTGATGACTTCCCAAAGCTCGATGAAGCCTGGCCCAAGAACGTGGACATCCGCTCGATCACGCCGGTGCTCGACACGGACACGGATTCGTGTTTGCCAAGCGCCATGGTGAGTCGCACTGCTCAGCAGAACGGAGGTCTCAAGCCAAGCGGTCAGATCGTCGGCAAGTGCTACAACCACGACTTTATGAATCTCTCCAACTCGTACCACCGCTACGCGTGCCAGAACAGTAATGGAAGCCAGTACTGCGCGCATTTGTTCGCGTACTACTTCCTCAAGGACCAGAAGATCAACTTCATCGAGAGTGGACATCGTCACGACTTGGAGCAGGTCGCAGTCTGGACAATCAACGGACGTGTCTCTCATGGCAGCTACAGTGCCCACGGCAAGATGTACGCCAAGGCGGCGAGTGACATCCCGAACCAGGACGGCCACCTCAAGTTCGTGTACCACAAGGACGGCACGGGTACCCACGCGTTCCGCTTTGCCAAGAACAACGAAGACGCGGAGAACCCACTGCGGACGTTCGTGTTCCCGACGGTTGTGAGCTGGTACACGATGAAGGGCGACGGCATCGACAATGCTGGTCTTCGAGACAAGATGAACAGTCACAACTACGGCAGCGCCACGATGCCGATCAAGGACAGCACGTTCCTCAACAACATCAACAACTTTCGTCCGTCGGGTTTCCCCGTCTTCACGCAGGACAGTGTCAACAACTCGCAG

PyolNLP2蛋白(261)

MKLLTILSVVCAAVAGSVRADDFPKLDEAWPKNVDIRSITPVLDTDTDSCLPSAMVSRTAQQNGGLKPSGQIVGKCYNHDFMNLSNSYHRYACQNSNGSQYCAHLFAYYFLKDQKINFIESGHRHDLEQVAVWTINGRVSHGSYSAHGKMYAKAASDIPNQDGHLKFVYHKDGTGTHAFRFAKNNEDAENPLRTFVFPTVVSWYTMKGDGIDNAGLRDKMNSHNYGSATMPIKDSTFLNNINNFRPSGFPVFTQDSVNNSQ

PyolNLP3基因(786bp)

ATGAACTGGTCCGCCATGCTCTCGCTCTGCGTCGCCACCGCCCTCAGTTCCGTCCACGCCGATGACTTCCCCAAGCTCGACGAGGCCATGCCTCGTAACATCGACATCCGTAGCATCACGCCCATGATCGACACGGACACAGACTCGTGTTTGCCGAGCGCTATGGTCAGCCGCACGGGTGCGCAGAACGCCGGATTGGACCCTGAGTGGGCCAGTTTGGTCAAGGGCTGTCGCAACAACGACTTCATGAACCTCTCCAACACGTACCATCGCTACGTGTGCCAGAACAACAACGGCAACCAGTACTGCGCACACTTGTTCGCGTACTACTTCCTCAAGGACCAGTCAGTGAATCTCATTCCAAGTGGTCACAAGAACGACTTGGAGCAGGTTGCGGTGTGGACGATCAACGGTCGTGTGTCGCATGGCAGCTACAGTGCTCACGGCAAGATGTACAACAAGGCCGCGAGCGAGATTCCAACCCAGGACGGTCATCTCAAGTTTGTGTACCACAAGGACGGTGGTAGTACGCACTGCTTCCGCTTCGCTGGTAACAACGAGTTCGCCGAGAACCCCGCGGGTACCTTTGTGACGCCAACGGTTGTGAGTTGGTACACGATGAAGGGCGACGGTATTGATAACGGTGGCCTTCGCGACCGTATGAACAACTACAACTACGGTAGTGCCACGATGCCGATCAAGGACAGCACGTTCCTTAACAACGTCAACACGTACAAGCCAGGCGAGTTCCCAGTCTTCACCCAGGACAGCGTGAACAACTCCCAG

PyolNLP3蛋白(262)

MNWSAMLSLCVATALSSVHADDFPKLDEAMPRNIDIRSITPMIDTDTDSCLPSAMVSRTGAQNAGLDPEWASLVKGCRNNDFMNLSNTYHRYVCQNNNGNQYCAHLFAYYFLKDQSVNLIPSGHKNDLEQVAVWTINGRVSHGSYSAHGKMYNKAASEIPTQDGHLKFVYHKDGGSTHCFRFAGNNEFAENPAGTFVTPTVVSWYTMKGDGIDNGGLRDRMNNYNYGSATMPIKDSTFLNNVNTYKPGEFPVFTQDSVNNSQ

PyolNLP4基因(777bp)

ATGAAGTGGATTGCCTTGCTCGCTTCTATCGCCTTGCTCGGCCGTGTTCGTGGTGATGATTTCCCCAAGCTCGATGCCGCGCCACCCAAGAACATCGATATCTTCTCCATCACGCCAGTTCTCGACATGGACACGGACTCGTGTCTTCCAAGTGCCGCCATCAGTCGTACTGGGGCCCAGAACGGAGGTCTCAACCCAAGCGGAAGTATCACTGGCGGCTGCCGACGCTCGGATTTCATGAACTATTCCAACACGTACCACCGTTACGCGTGTATCAGCAGCAACGGTAACCAGTACTGCGGGCACTTCTTCGCCTACTACTTCCTCAAGGACCAGGTATTGAGCACGGTTGGTGGTGGCCACCGTCACGATATTGAGCAGGTTGGTATCTGGACAACCAACGGCCGAGTCACCCACGGTGGCTTCAGCGCCCATGGGGCCATGACGAATCTCCCGGCTAACCAGATCCCTCAAGAAGGTGGTCGCCTCAAATTCGTGTACCACAAGGACGGAGTGGCAACGCACGCGATCCGCTTTGCCAAGACTAATGAGGCTGTTGAGAACCCAACAGGAGCCTTCGTCGTGCCTACACTGGTGAGCTGGTTCACGATGAAGGCCGACAGTATCAACAACGCTGGACTGCGCAACAACTTGAACTCGTTTGACTACGGCAACGCCAACATGCCGATCAAGGACAGCAACTTCCTGAACAGCTTGAATAAGTTCCGTCCAGCCGAGTATCCTGCGTTTACACAGGCCAGCGTCGACAACTCTCAG

PyolNLP4蛋白(259)

MKWIALLASIALLGRVRGDDFPKLDAAPPKNIDIFSITPVLDMDTDSCLPSAAISRTGAQNGGLNPSGSITGGCRRSDFMNYSNTYHRYACISSNGNQYCGHFFAYYFLKDQVLSTVGGGHRHDIEQVGIWTTNGRVTHGGFSAHGAMTNLPANQIPQEGGRLKFVYHKDGVATHAIRFAKTNEAVENPTGAFVVPTLVSWFTMKADSINNAGLRNNLNSFDYGNANMPIKDSNFLNSLNKFRPAEYPAFTQASVDNSQ

PyolNLP5基因(786bp)

ATGAACTGGTCCGCCATCCTCTCGCTCTGTGTCGCCGCCGCCCTCGGTTCCGTCCACGCCGATGACTTCCCCAAGCTCGACGAGGCCATGCCTCGTAACATCGACATTCGTGCCATCACGCCCGTGGTTGACACGGATACCGATGGCTGTCTCCCAAGCGCCATGGTGAGCCGCACCGGTCAGCAGAACGGTGGTTTGGCTCCTGGTTGGAGCAACCTCACCAAGGGCTGTCGCAACAACGACTTCATGAACCTCTCCAACACGTACCATCGTTACGTGTGCCAGAACAACAACGGCAACCAGTACTGCGCGCACTTGTTCGCGTACTACTTCCTCAAGGATCAGTCGGTCAACCTCATTCCAAGTGGTCACAAGCACGACTTGGAGCAGGTTGCTGTTTGGACGATCAACGGTCGTGTGTCGCATGGCAGCTACAGTGCTCACGGCAAGATGTACAGCAAGCCAGCAAGCGAGATCCCAACCCAGGACGGCCACCTCAAGTTTGTGTACCACAAGGACGGTGGTAGCACGCACTGCTTCCGCTTCGCTGGTAACAACGAGTTCGCCGAGAACCCCGCGGGTACTTTCGTCACTCCTACTCTCGTGAGTTGGTACACGATGAAGGGCGACGGTATTGATAACGGTGGCCTTCGTGACAAGATGAACAACTACAACTACGGCAGCGCTTCGATGCCGATCAAGGACGGCTCGTTCCTTAACAACGTCAACACGTACAAGCCAGGCGAGTTCCCAACGTTCACCCAGGACAGTGTGAACAACTCCCAG

PyolNLP5蛋白(262)

MNWSAILSLCVAAALGSVHADDFPKLDEAMPRNIDIRAITPVVDTDTDGCLPSAMVSRTGQQNGGLAPGWSNLTKGCRNNDFMNLSNTYHRYVCQNNNGNQYCAHLFAYYFLKDQSVNLIPSGHKHDLEQVAVWTINGRVSHGSYSAHGKMYSKPASEIPTQDGHLKFVYHKDGGSTHCFRFAGNNEFAENPAGTFVTPTLVSWYTMKGDGIDNGGLRDKMNNYNYGSASMPIKDGSFLNNVNTYKPGEFPTFTQDSVNNSQ

PyolNLP6基因(786bp)

ATGAAGTGGTCCGCCATCCTCTCGCTCTGTGTCGCCGCCGTCCTAGGTTCGGCCCACGCCGATGACTTCCCCAAGCTCGACGAGGCCATGCCCAAGAACGTCGACATCCGCGGTATTACGCCCGTGATCGACACGGATATGGATTCGTGTCTTCCTAGTGCCATGGTCAGTCGTACTGGTCAGCAGAACGGCGGGTTGGAACCAGCGTGGGCCAGTTTGGTCAAGGGATGTCGCAACAACGACTTCATGAAGCTCTCCAACACGTACCATCGTTACGTATGCCAGACGAGTAACAACAACCAGTACTGTGCGCACTTGTTTGCGTACTACTTCCTCAAGGATCAGTCGGTCAACCTGATTCCAAGCGGTCATAAGCATGACCTGGAATATGTCGCAGTGTGGACAATTAATGGACGCATGACGCATGGTAGCTACAGCGCTCATGGCAAGACCTACAACAAGCCAGCGACCGAAATTCCAACCCAAGACGGTCATCTCAAGTTCGTGTATCACAAGGACGGTGGCTTGACGCACTGCTTTAGATTCGCGCAGAAGAACGAGAACGCAGAGAATCCAGCGGGTACCTTCGTCTTGCCGACGCTTGCGAGCTGGTATACGATGAAAGGCGATGGTATCGACAATGCCGGTCTTCGTAACAAGTTGAACAGCTACAGCTATGGGAATGCGAACATGCCGCTCAAGGACGGCAAGTTCCTCGAC

AAGGTCAACTCGTTCAAGCCATCCGAGTTTCCAAGCTTCACTGAGGACCATGCAAAGAACTCTCAG

PyolNLP6蛋白(262)

MKWSAILSLCVAAVLGSAHADDFPKLDEAMPKNVDIRGITPVIDTDMDSCLPSAMVSRTGQQNGGLEPAWASLVKGCRNNDFMKLSNTYHRYVCQTSNNNQYCAHLFAYYFLKDQSVNLIPSGHKHDLEYVAVWTINGRMTHGSYSAHGKTYNKPATEIPTQDGHLKFVYHKDGGLTHCFRFAQKNENAENPAGTFVLPTLASWYTMKGDGIDNAGLRNKLNSYSYGNANMPLKDGKFLDKVNSFKPSEFPSFTEDHAKNSQ

PyolNLP7基因(780bp)

ATGAAGTGGGCCGCGCTCCTTGTCTTCTCCGCCGCCTTGCTCTCCACTGTCCGCGGCGACGACTTCCCCAAGCTCGATGCTGCACCACCAAAGAACATTGACATCTCGTCCATTACGCCAGTTCTTGACATGGACACGGACTCGTGTCTTCCAAGTGCGGCAATCAGTCGGAGTGGACAGCAGAATGGCGGCCTCAAGCCAACTGGAAAGATTACTGGTGGTTGTCGTCGCGGGGACTTCATGGACTTCTCAAACACGTACCATCGCTACGCGTGTGTGAACGGCAACGGCAACCAGTACTGTGGCCACTTCTTCGCTTACTACTTCCTCAAGGATCAAGCCGTCCCCACGTTTGGCGGTGGCCATCGCCACGACCTCGAGCAGATCGGTATCTGGACCGTCAATGGCCGAGTCACCCACGGTGGCTTCAGTGCCCATGGCAAGATGGAGAACCGCCCAGTGGACCAGATTCCGCAGGAAGGTGGTCGCCTCAAGTTTGTCTACCACAAGGACGGAGTGGTGACGCACGCGATCCGCTTTGGCAAGACCAACGAAGGCGCCGAGAACCCAACGGGAGCCTTCGTTGTGCCCACATTGGTGAGCTGGTTCACGATGAAGGGCGATACCATCGACAACGCTGGACTGCGTAACAACTTGAACTCGTTTGACTACGGCAACGCCAACATGCCGATCAAGGACAACAACTTCTTGAGCAACTTGAACAAGTTTCGTCCATCCGACTTTCCAGAGTTTACGCAAGAGAGTATCGACACCTCCCAG

PyolNLP7蛋白(260)

MKWAALLVFSAALLSTVRGDDFPKLDAAPPKNIDISSITPVLDMDTDSCLPSAAISRSGQQNGGLKPTGKITGGCRRGDFMDFSNTYHRYACVNGNGNQYCGHFFAYYFLKDQAVPTFGGGHRHDLEQIGIWTVNGRVTHGGFSAHGKMENRPVDQIPQEGGRLKFVYHKDGVVTHAIRFGKTNEGAENPTGAFVVPTLVSWFTMKGDTIDNAGLRNNLNSFDYGNANMPIKDNNFLSNLNKFRPSDFPEFTQESIDTSQ

PyolNLP9基因(795bp)

ATGAAACCAGCCACATTCCTCACTTTTCTCGCTATCGGTCTCCAGGCCACCGCCACCTACGCCCACGACTTCCCCAAGATGGACGAAGCCTTGCCCAAGAACATCGACATCCGCTCGATTACTCCTCTGCTTGACTTTGATGAGGACTCGTGCCTCCCGAGCACGGCTGTAAGCCGCGAGGGTAAGCAGAACGGTGGTCTCAAGACGACTGGTAACATCGCCGGAGGCTGTCGGGATAAGGAAGACTTCCTGGAGTACTCCAACACTTACTATCGCTACGTGTGCGCCAACTACGAAGGGAAGAAATACTGTGGTCACATGTTTGCGCTCTACTTCCAGAAGGACCAGGCGACTCTTCTTGGCTTGGGCCATCGTCACGACTGGGAACATGTGACAGTGTGGACCACTGATGGTATTGTCACTCACGGCAGCGCGAGTGCTCACGGAGGTGGCAAGACTGCGTCCTTTGGTGCCATCCCGAATGAGGACCAGCACCTTCTCTTCGTGTACCACAAGGACGACGTACAGACGCATGCCATGCGCTTTGCTGATGCTGATGACCAGGCGGAGGCAGAAAACCCATACAAGCAGTTCGTCTTGCCTCCTGTTGTGAGCTGGTACAGCATGCAGGGCGACGGTGCCCTGGACAACGAAACACTGCGCAATATGATGAACGGGCTAAACTTTGGTAGCGCTCACTGTCCGCTTCTGGACGGCGATTTCCTGCGCAACCTAAACAGCCTCAAGCCAGAGGGGTACCCGACGTTCACAGACGCTGACATGATCCAATCGAAG

PyolNLP9蛋白(265)

MKPATFLTFLAIGLQATATYAHDFPKMDEALPKNIDIRSITPLLDFDEDSCLPSTAVSREGKQNGGLKTTGNIAGGCRDKEDFLEYSNTYYRYVCANYEGKKYCGHMFALYFQKDQATLLGLGHRHDWEHVTVWTTDGIVTHGSASAHGGGKTASFGAIPNEDQHLLFVYHKDDVQTHAMRFADADDQAEAENPYKQFVLPPVVSWYSMQGDGALDNETLRNMMNGLNFGSAHCPLLDGDFLRNLNSLKPEGYPTFTDADMIQSK

PypeNLP4基因(298bp)

ATGAAGTTCGCCTCGATTCTCGCTTTCCTCACCGTCGGAATTCAAGTGAGTATCACACAAGCCCACGACTTTCCCCGATTAGATGAAGCCATGCCAAAGGGCATCGACATCAAGGCCATCTTGCCACTACTTGACTTCGACGACGACTCGTGCTATCCCAGCGCGGCTGTGAGTCGAGTGGGTCGCCAGAACGGTGGTCTTCGGACCACAGGTTCCATCACAGGCGGTTGCCGCAACAAAGAGTTCCTCGACTACTCAAACTCGTACCACCGCTATGCGTGTCGAGACCACGAGAAGAAGAAATACTGCGGCCACATGTTTGCTCTGTATTTCCAGAAGGATCAAATGGTCGACGGACTGGACCCTTTCGGACACCGTCACGACTTTGAATATGTCGCTGTGTGGACAACCGACGGTGTCGTCACGCATGGCAGCGCCAGTGCCCACGGTGACGCGGCAGTGAAGCCTATCGATCAGATCCCACACTCGGGCGAGCATATCAAGTTCGTATTCCACAAGGACGAACCGACAACGCACTCGATGCGTTTCGCTGAGAAGAACCACGACGAAGACGCCGAGAACTCGTACGGCGACTTTGTCCTGCCTGCCGTTGTGAGCTGGTACACTATGCAGGGCGACGGCAACATGACCAATGATGCCTTGCGTGGCCTGATGAACAGTCTCGACTTTGGCGACGCCCACAACCCGCTTCTGGATGGTGATTTCTTGGATAATCTGAACGAAGCCAAGCCGAAGGAGTACCCTCAGTTTGACGATGAGGATGTCAAGGGCTCGCAG

PypeNLP4蛋白(266)

MKFASILAFLTVGIQVSITQAHDFPRLDEAMPKGIDIKAILPLLDFDDDSCYPSAAVSRVGRQNGGLRTTGSITGGCRNKEFLDYSNSYHRYACRDHEKKKYCGHMFALYFQKDQMVDGLDPFGHRHDFEYVAVWTTDGVVTHGSASAHGDAAVKPIDQIPHSGEHIKFVFHKDEPTTHSMRFAEKNHDEDAENSYGDFVLPAVVSWYTMQGDGNMTNDALRGLMNSLDFGDAHNPLLDGDFLDNLNEAKPKEYPQFDDEDVKGSQ

PypeNLP5基因(795bp)

ATGAAACCAGCCACATTCCTCACTTTTCTCGCTATCGGTCTCCAGGCCACCGCCACCTACGCCCACGACTTCCCCAAGATGGACGAAGCCTTGCCCAAGAACATCGACATCCGCTCGATTACTCCTCTGCTTGACTTTGATGAGGACTCGTGCCTCCCGAGCACGGCTGTAAGCCGCGAGGGTAAGCAGAACGGTGGTCTCAAGACGAGTGGTAACATCGCTGGAGGCTGTCGGGATAAGGAAGACTTCCTGGAGTACTCCAACACTTACCATCGCTACGTGTGCGCCAACTACGAAGGGAAGAAATACTGTGGTCACATGTTTGCGCTCTACTTCCAGAAGGACCAGGCGACTCTTCTTGGCTTGGGCCATCGTCACGACTGGGAGCATGTGACAGTGTGGACCACTGATGGTATTGTCACTCACGGCAGCGCGAGTGCTCACGGAGGTGGCAAGACTGCGTCCTTTGATGCCATCCCGAATGAGGACCAGCACCTTCTCTTCGTGTACCACAAGGACGACGTACAGACGCACGCCATGCGCTTTGCTGATGCTGATGACCAGGCGGAGGTAGAAAACCCATACCTGCAGTTCGTCTTGCCTCCTGTTGTGAGCTGGTACAGCATGCAGGGCGACGGTGCCCTGGACAACGAAACACTGCGCAATATGATGAACAGGCTAAACTTTGGTAGCGCTCACTGTCCGCTTCTGGACGGCGATTTCCTGCGCAACCTAAACAGCCTCAAGCCAGAGGGGTACCCGACGTTCACGGACGCTGACATGATCCAATCGAAG

PypeNLP5蛋白(265)

MKPATFLTFLAIGLQATATYAHDFPKMDEALPKNIDIRSITPLLDFDEDSCLPSTAVSREGKQNGGLKTSGNIAGGCRDKEDFLEYSNTYHRYVCANYEGKKYCGHMFALYFQKDQATLLGLGHRHDWEHVTVWTTDGIVTHGSASAHGGGKTASFDAIPNEDQHLLFVYHKDDVQTHAMRFADADDQAEVENPYLQFVLPPVVSWYSMQGDGALDNETLRNMMNRLNFGSAHCPLLDGDFLRNLNSLKPEGYPTFTDADMIQSK

PypeNLP8基因(783bp)

ATGAAGCTTTTGACGATCCTCTCCGTCGTATGCGCTGCTGTCGCTGGCTCTGTGCGCGCTGATGACTTCCCAAAGCTCGATGAAGCCTGGCCCAAGAACGTGGACATCCGCTCGATCACGCCGGTGTTTGATATGGACACGGATTCGTGTTTGCCAAGCGCCATGGTGAGTCGCACTGGTCAGCAGAACGGAGGTCTCAAGCCAAGCGGTCAGATCGTCGGCAAGTGCTACAACCACGACTTTATGAATCTCTCCAACTCGTACCACCGCTACGCTTGCCAGAACAGTAATGGAAGCCAGTACTGCGCGCATTTGTTCGCGTACTACTTCCTCAAGGACCAGAAGATCAACTTCATCGAGAGTGGACATCGTCACGACTTGGAGCAGCTCGCAGTCTGGACGATCAACGGACGTGTCTCTCATGGCAGCTACAGTGCCCACGGCAAGATGTACGCCAAGGCGGCGAGTGACATCCCGAACCAGGACGGCCACCTCAAGTTCGTGTACCACAAGGACGGCGCGGGTACCCACGCGTTCCGCTTTGCCAAGAACAACGAAGACGCGGAGAACCCACTGCGGACGTTCGTGTTCCCGACGGTTGTGAGCTGGTACACGATGAAGGGCGACGGCATCGACAATGCTGGTCTTCGAGACAAGATGAACAGCTACAACTACGGCAGCGCCACGATGCCGATCAAGGACAGCACGTTCCTCAACAACATCAACAACTTTCGTCCGTCTGGTTTCCCCGTCTTCACGCAGGACAGTGTCAACAACTCGCAG

PypeNLP8蛋白(261)

MKLLTILSVVCAAVAGSVRADDFPKLDEAWPKNVDIRSITPVFDMDTDSCLPSAMVSRTGQQNGGLKPSGQIVGKCYNHDFMNLSNSYHRYACQNSNGSQYCAHLFAYYFLKDQKINFIESGHRHDLEQLAVWTINGRVSHGSYSAHGKMYAKAASDIPNQDGHLKFVYHKDGAGTHAFRFAKNNEDAENPLRTFVFPTVVSWYTMKGDGIDNAGLRDKMNSYNYGSATMPIKDSTFLNNINNFRPSGFPVFTQDSVNNSQ

序列表

<110> 南京农业大学

<120> 一种生防腐霉菌中诱导坏死和活性氧积累的NLP类基因及其表达载体和应用

<160> 22

<170> SIPOSequenceListing 1.0

<210> 1

<211> 810

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 1

atggcagtcc gttgcttggt ctctacgatc gtgcttgcta ttgtgccact cgccagtggg 60

attggtttcg aggtaatcac aggtgacgtc ttcccgagtc tctcgcgctc atggcccaag 120

cacgtggaca tctcgtcgat tacgccactc tttgacatgg atgacaatgg ctgctaccca 180

gcggccgtca ttagtcgctc tggtgagccg aaccgaggcc tggagaccga cggcggcaac 240

tatgatgtct gtaaacgcga cgatttcctc gagtactcga atacgtatca ccgctacgcg 300

tgtcgtgtcg ccgataacat caagtactgc agtcatttct tcacgctcta cttccaaaaa 360

gacgaagcaa ctctcatcgg tggccatcgt cacgacctcg agcaagtcgt catctggacc 420

atggacggca acgtcactca cgggagcttc agcgcccatg gtggtatgac cacctggtca 480

gcgagcagca tcccacacga aggctcgcat ctcaaattcg tgtatcacaa agacaacatc 540

atgacgcacg ccatgcgctt tgcaaggacc aaagacgcgt tggacccaga gaatccgtac 600

aagaagtttg tgcttccacc agtcgtgagc tggtacacga tgaagggcga tggagatctg 660

accaacagtg cgttgcgtca gcttctcaac acacttcctt acggcaaagc gactatgccc 720

atcaaggact ccgtatttct gaggaacctc aatatgagac gaccccaagg ataccccgag 780

tttgacgatg aagacatcaa gagctcgcaa 810

<210> 2

<211> 783

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 2

atgaagcttt tgacgatcct ctccgtcgta tgcgctgctg tcgctggctc tgtgcgcgct 60

gatgacttcc caaagctcga tgaagcctgg cccaagaacg tggacatccg ctcgatcacg 120

ccggtgctcg acacggacac ggattcgtgt ttgccaagcg ccatggtgag tcgcactgct 180

cagcagaacg gaggtctcaa gccaagcggt cagatcgtcg gcaagtgcta caaccacgac 240

tttatgaatc tctccaactc gtaccaccgc tacgcgtgcc agaacagtaa tggaagccag 300

tactgcgcgc atttgttcgc gtactacttc ctcaaggacc agaagatcaa cttcatcgag 360

agtggacatc gtcacgactt ggagcaggtc gcagtctgga caatcaacgg acgtgtctct 420

catggcagct acagtgccca cggcaagatg tacgccaagg cggcgagtga catcccgaac 480

caggacggcc acctcaagtt cgtgtaccac aaggacggca cgggtaccca cgcgttccgc 540

tttgccaaga acaacgaaga cgcggagaac ccactgcgga cgttcgtgtt cccgacggtt 600

gtgagctggt acacgatgaa gggcgacggc atcgacaatg ctggtcttcg agacaagatg 660

aacagtcaca actacggcag cgccacgatg ccgatcaagg acagcacgtt cctcaacaac 720

atcaacaact ttcgtccgtc gggtttcccc gtcttcacgc aggacagtgt caacaactcg 780

cag 783

<210> 3

<211> 786

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 3

atgaactggt ccgccatgct ctcgctctgc gtcgccaccg ccctcagttc cgtccacgcc 60

gatgacttcc ccaagctcga cgaggccatg cctcgtaaca tcgacatccg tagcatcacg 120

cccatgatcg acacggacac agactcgtgt ttgccgagcg ctatggtcag ccgcacgggt 180

gcgcagaacg ccggattgga ccctgagtgg gccagtttgg tcaagggctg tcgcaacaac 240

gacttcatga acctctccaa cacgtaccat cgctacgtgt gccagaacaa caacggcaac 300

cagtactgcg cacacttgtt cgcgtactac ttcctcaagg accagtcagt gaatctcatt 360

ccaagtggtc acaagaacga cttggagcag gttgcggtgt ggacgatcaa cggtcgtgtg 420

tcgcatggca gctacagtgc tcacggcaag atgtacaaca aggccgcgag cgagattcca 480

acccaggacg gtcatctcaa gtttgtgtac cacaaggacg gtggtagtac gcactgcttc 540

cgcttcgctg gtaacaacga gttcgccgag aaccccgcgg gtacctttgt gacgccaacg 600

gttgtgagtt ggtacacgat gaagggcgac ggtattgata acggtggcct tcgcgaccgt 660

atgaacaact acaactacgg tagtgccacg atgccgatca aggacagcac gttccttaac 720

aacgtcaaca cgtacaagcc aggcgagttc ccagtcttca cccaggacag cgtgaacaac 780

tcccag 786

<210> 4

<211> 777

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 4

atgaagtgga ttgccttgct cgcttctatc gccttgctcg gccgtgttcg tggtgatgat 60

ttccccaagc tcgatgccgc gccacccaag aacatcgata tcttctccat cacgccagtt 120

ctcgacatgg acacggactc gtgtcttcca agtgccgcca tcagtcgtac tggggcccag 180

aacggaggtc tcaacccaag cggaagtatc actggcggct gccgacgctc ggatttcatg 240

aactattcca acacgtacca ccgttacgcg tgtatcagca gcaacggtaa ccagtactgc 300

gggcacttct tcgcctacta cttcctcaag gaccaggtat tgagcacggt tggtggtggc 360

caccgtcacg atattgagca ggttggtatc tggacaacca acggccgagt cacccacggt 420

ggcttcagcg cccatggggc catgacgaat ctcccggcta accagatccc tcaagaaggt 480

ggtcgcctca aattcgtgta ccacaaggac ggagtggcaa cgcacgcgat ccgctttgcc 540

aagactaatg aggctgttga gaacccaaca ggagccttcg tcgtgcctac actggtgagc 600

tggttcacga tgaaggccga cagtatcaac aacgctggac tgcgcaacaa cttgaactcg 660

tttgactacg gcaacgccaa catgccgatc aaggacagca acttcctgaa cagcttgaat 720

aagttccgtc cagccgagta tcctgcgttt acacaggcca gcgtcgacaa ctctcag 777

<210> 5

<211> 786

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 5

atgaactggt ccgccatcct ctcgctctgt gtcgccgccg ccctcggttc cgtccacgcc 60

gatgacttcc ccaagctcga cgaggccatg cctcgtaaca tcgacattcg tgccatcacg 120

cccgtggttg acacggatac cgatggctgt ctcccaagcg ccatggtgag ccgcaccggt 180

cagcagaacg gtggtttggc tcctggttgg agcaacctca ccaagggctg tcgcaacaac 240

gacttcatga acctctccaa cacgtaccat cgttacgtgt gccagaacaa caacggcaac 300

cagtactgcg cgcacttgtt cgcgtactac ttcctcaagg atcagtcggt caacctcatt 360

ccaagtggtc acaagcacga cttggagcag gttgctgttt ggacgatcaa cggtcgtgtg 420

tcgcatggca gctacagtgc tcacggcaag atgtacagca agccagcaag cgagatccca 480

acccaggacg gccacctcaa gtttgtgtac cacaaggacg gtggtagcac gcactgcttc 540

cgcttcgctg gtaacaacga gttcgccgag aaccccgcgg gtactttcgt cactcctact 600

ctcgtgagtt ggtacacgat gaagggcgac ggtattgata acggtggcct tcgtgacaag 660

atgaacaact acaactacgg cagcgcttcg atgccgatca aggacggctc gttccttaac 720

aacgtcaaca cgtacaagcc aggcgagttc ccaacgttca cccaggacag tgtgaacaac 780

tcccag 786

<210> 6

<211> 786

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 6

atgaagtggt ccgccatcct ctcgctctgt gtcgccgccg tcctaggttc ggcccacgcc 60

gatgacttcc ccaagctcga cgaggccatg cccaagaacg tcgacatccg cggtattacg 120

cccgtgatcg acacggatat ggattcgtgt cttcctagtg ccatggtcag tcgtactggt 180

cagcagaacg gcgggttgga accagcgtgg gccagtttgg tcaagggatg tcgcaacaac 240

gacttcatga agctctccaa cacgtaccat cgttacgtat gccagacgag taacaacaac 300

cagtactgtg cgcacttgtt tgcgtactac ttcctcaagg atcagtcggt caacctgatt 360

ccaagcggtc ataagcatga cctggaatat gtcgcagtgt ggacaattaa tggacgcatg 420

acgcatggta gctacagcgc tcatggcaag acctacaaca agccagcgac cgaaattcca 480

acccaagacg gtcatctcaa gttcgtgtat cacaaggacg gtggcttgac gcactgcttt 540

agattcgcgc agaagaacga gaacgcagag aatccagcgg gtaccttcgt cttgccgacg 600

cttgcgagct ggtatacgat gaaaggcgat ggtatcgaca atgccggtct tcgtaacaag 660

ttgaacagct acagctatgg gaatgcgaac atgccgctca aggacggcaa gttcctcgac 720

aaggtcaact cgttcaagcc atccgagttt ccaagcttca ctgaggacca tgcaaagaac 780

tctcag 786

<210> 7

<211> 780

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 7

atgaagtggg ccgcgctcct tgtcttctcc gccgccttgc tctccactgt ccgcggcgac 60

gacttcccca agctcgatgc tgcaccacca aagaacattg acatctcgtc cattacgcca 120

gttcttgaca tggacacgga ctcgtgtctt ccaagtgcgg caatcagtcg gagtggacag 180

cagaatggcg gcctcaagcc aactggaaag attactggtg gttgtcgtcg cggggacttc 240

atggacttct caaacacgta ccatcgctac gcgtgtgtga acggcaacgg caaccagtac 300

tgtggccact tcttcgctta ctacttcctc aaggatcaag ccgtccccac gtttggcggt 360

ggccatcgcc acgacctcga gcagatcggt atctggaccg tcaatggccg agtcacccac 420

ggtggcttca gtgcccatgg caagatggag aaccgcccag tggaccagat tccgcaggaa 480

ggtggtcgcc tcaagtttgt ctaccacaag gacggagtgg tgacgcacgc gatccgcttt 540

ggcaagacca acgaaggcgc cgagaaccca acgggagcct tcgttgtgcc cacattggtg 600

agctggttca cgatgaaggg cgataccatc gacaacgctg gactgcgtaa caacttgaac 660

tcgtttgact acggcaacgc caacatgccg atcaaggaca acaacttctt gagcaacttg 720

aacaagtttc gtccatccga ctttccagag tttacgcaag agagtatcga cacctcccag 780

<210> 8

<211> 795

<212> DNA

<213> 寡雄腐霉(Pythium oligandrum)

<400> 8

atgaaaccag ccacattcct cacttttctc gctatcggtc tccaggccac cgccacctac 60

gcccacgact tccccaagat ggacgaagcc ttgcccaaga acatcgacat ccgctcgatt 120

actcctctgc ttgactttga tgaggactcg tgcctcccga gcacggctgt aagccgcgag 180

ggtaagcaga acggtggtct caagacgact ggtaacatcg ccggaggctg tcgggataag 240

gaagacttcc tggagtactc caacacttac tatcgctacg tgtgcgccaa ctacgaaggg 300

aagaaatact gtggtcacat gtttgcgctc tacttccaga aggaccaggc gactcttctt 360

ggcttgggcc atcgtcacga ctgggaacat gtgacagtgt ggaccactga tggtattgtc 420

actcacggca gcgcgagtgc tcacggaggt ggcaagactg cgtcctttgg tgccatcccg 480

aatgaggacc agcaccttct cttcgtgtac cacaaggacg acgtacagac gcatgccatg 540

cgctttgctg atgctgatga ccaggcggag gcagaaaacc catacaagca gttcgtcttg 600

cctcctgttg tgagctggta cagcatgcag ggcgacggtg ccctggacaa cgaaacactg 660

cgcaatatga tgaacgggct aaactttggt agcgctcact gtccgcttct ggacggcgat 720

ttcctgcgca acctaaacag cctcaagcca gaggggtacc cgacgttcac agacgctgac 780

atgatccaat cgaag 795

<210> 9

<211> 798

<212> DNA

<213> 缠器腐霉(Pythium periplocum)

<400> 9

atgaagttcg cctcgattct cgctttcctc accgtcggaa ttcaagtgag tatcacacaa 60

gcccacgact ttccccgatt agatgaagcc atgccaaagg gcatcgacat caaggccatc 120

ttgccactac ttgacttcga cgacgactcg tgctatccca gcgcggctgt gagtcgagtg 180

ggtcgccaga acggtggtct tcggaccaca ggttccatca caggcggttg ccgcaacaaa 240

gagttcctcg actactcaaa ctcgtaccac cgctatgcgt gtcgagacca cgagaagaag 300

aaatactgcg gccacatgtt tgctctgtat ttccagaagg atcaaatggt cgacggactg 360

gaccctttcg gacaccgtca cgactttgaa tatgtcgctg tgtggacaac cgacggtgtc 420

gtcacgcatg gcagcgccag tgcccacggt gacgcggcag tgaagcctat cgatcagatc 480

ccacactcgg gcgagcatat caagttcgta ttccacaagg acgaaccgac aacgcactcg 540

atgcgtttcg ctgagaagaa ccacgacgaa gacgccgaga actcgtacgg cgactttgtc 600

ctgcctgccg ttgtgagctg gtacactatg cagggcgacg gcaacatgac caatgatgcc 660

ttgcgtggcc tgatgaacag tctcgacttt ggcgacgccc acaacccgct tctggatggt 720

gatttcttgg ataatctgaa cgaagccaag ccgaaggagt accctcagtt tgacgatgag 780

gatgtcaagg gctcgcag 798

<210> 10

<211> 795

<212> DNA

<213> 缠器腐霉(Pythium periplocum)

<400> 10

atgaaaccag ccacattcct cacttttctc gctatcggtc tccaggccac cgccacctac 60

gcccacgact tccccaagat ggacgaagcc ttgcccaaga acatcgacat ccgctcgatt 120

actcctctgc ttgactttga tgaggactcg tgcctcccga gcacggctgt aagccgcgag 180

ggtaagcaga acggtggtct caagacgagt ggtaacatcg ctggaggctg tcgggataag 240

gaagacttcc tggagtactc caacacttac catcgctacg tgtgcgccaa ctacgaaggg 300

aagaaatact gtggtcacat gtttgcgctc tacttccaga aggaccaggc gactcttctt 360

ggcttgggcc atcgtcacga ctgggagcat gtgacagtgt ggaccactga tggtattgtc 420

actcacggca gcgcgagtgc tcacggaggt ggcaagactg cgtcctttga tgccatcccg 480

aatgaggacc agcaccttct cttcgtgtac cacaaggacg acgtacagac gcacgccatg 540

cgctttgctg atgctgatga ccaggcggag gtagaaaacc catacctgca gttcgtcttg 600

cctcctgttg tgagctggta cagcatgcag ggcgacggtg ccctggacaa cgaaacactg 660

cgcaatatga tgaacaggct aaactttggt agcgctcact gtccgcttct ggacggcgat 720

ttcctgcgca acctaaacag cctcaagcca gaggggtacc cgacgttcac ggacgctgac 780

atgatccaat cgaag 795

<210> 11

<211> 783

<212> DNA

<213> 缠器腐霉(Pythium periplocum)

<400> 11

atgaagcttt tgacgatcct ctccgtcgta tgcgctgctg tcgctggctc tgtgcgcgct 60

gatgacttcc caaagctcga tgaagcctgg cccaagaacg tggacatccg ctcgatcacg 120

ccggtgtttg atatggacac ggattcgtgt ttgccaagcg ccatggtgag tcgcactggt 180

cagcagaacg gaggtctcaa gccaagcggt cagatcgtcg gcaagtgcta caaccacgac 240

tttatgaatc tctccaactc gtaccaccgc tacgcttgcc agaacagtaa tggaagccag 300

tactgcgcgc atttgttcgc gtactacttc ctcaaggacc agaagatcaa cttcatcgag 360

agtggacatc gtcacgactt ggagcagctc gcagtctgga cgatcaacgg acgtgtctct 420

catggcagct acagtgccca cggcaagatg tacgccaagg cggcgagtga catcccgaac 480

caggacggcc acctcaagtt cgtgtaccac aaggacggcg cgggtaccca cgcgttccgc 540

tttgccaaga acaacgaaga cgcggagaac ccactgcgga cgttcgtgtt cccgacggtt 600

gtgagctggt acacgatgaa gggcgacggc atcgacaatg ctggtcttcg agacaagatg 660

aacagctaca actacggcag cgccacgatg ccgatcaagg acagcacgtt cctcaacaac 720

atcaacaact ttcgtccgtc tggtttcccc gtcttcacgc aggacagtgt caacaactcg 780

cag 783

<210> 12

<211> 270

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 12

Met Ala Val Arg Cys Leu Val Ser Thr Ile Val Leu Ala Ile Val Pro

1 5 10 15

Leu Ala Ser Gly Ile Gly Phe Glu Val Ile Thr Gly Asp Val Phe Pro

20 25 30

Ser Leu Ser Arg Ser Trp Pro Lys His Val Asp Ile Ser Ser Ile Thr

35 40 45

Pro Leu Phe Asp Met Asp Asp Asn Gly Cys Tyr Pro Ala Ala Val Ile

50 55 60

Ser Arg Ser Gly Glu Pro Asn Arg Gly Leu Glu Thr Asp Gly Gly Asn

65 70 75 80

Tyr Asp Val Cys Lys Arg Asp Asp Phe Leu Glu Tyr Ser Asn Thr Tyr

85 90 95

His Arg Tyr Ala Cys Arg Val Ala Asp Asn Ile Lys Tyr Cys Ser His

100 105 110

Phe Phe Thr Leu Tyr Phe Gln Lys Asp Glu Ala Thr Leu Ile Gly Gly

115 120 125

His Arg His Asp Leu Glu Gln Val Val Ile Trp Thr Met Asp Gly Asn

130 135 140

Val Thr His Gly Ser Phe Ser Ala His Gly Gly Met Thr Thr Trp Ser

145 150 155 160

Ala Ser Ser Ile Pro His Glu Gly Ser His Leu Lys Phe Val Tyr His

165 170 175

Lys Asp Asn Ile Met Thr His Ala Met Arg Phe Ala Arg Thr Lys Asp

180 185 190

Ala Leu Asp Pro Glu Asn Pro Tyr Lys Lys Phe Val Leu Pro Pro Val

195 200 205

Val Ser Trp Tyr Thr Met Lys Gly Asp Gly Asp Leu Thr Asn Ser Ala

210 215 220

Leu Arg Gln Leu Leu Asn Thr Leu Pro Tyr Gly Lys Ala Thr Met Pro

225 230 235 240

Ile Lys Asp Ser Val Phe Leu Arg Asn Leu Asn Met Arg Arg Pro Gln

245 250 255

Gly Tyr Pro Glu Phe Asp Asp Glu Asp Ile Lys Ser Ser Gln

260 265 270

<210> 13

<211> 261

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 13

Met Lys Leu Leu Thr Ile Leu Ser Val Val Cys Ala Ala Val Ala Gly

1 5 10 15

Ser Val Arg Ala Asp Asp Phe Pro Lys Leu Asp Glu Ala Trp Pro Lys

20 25 30

Asn Val Asp Ile Arg Ser Ile Thr Pro Val Leu Asp Thr Asp Thr Asp

35 40 45

Ser Cys Leu Pro Ser Ala Met Val Ser Arg Thr Ala Gln Gln Asn Gly

50 55 60

Gly Leu Lys Pro Ser Gly Gln Ile Val Gly Lys Cys Tyr Asn His Asp

65 70 75 80

Phe Met Asn Leu Ser Asn Ser Tyr His Arg Tyr Ala Cys Gln Asn Ser

85 90 95

Asn Gly Ser Gln Tyr Cys Ala His Leu Phe Ala Tyr Tyr Phe Leu Lys

100 105 110

Asp Gln Lys Ile Asn Phe Ile Glu Ser Gly His Arg His Asp Leu Glu

115 120 125

Gln Val Ala Val Trp Thr Ile Asn Gly Arg Val Ser His Gly Ser Tyr

130 135 140

Ser Ala His Gly Lys Met Tyr Ala Lys Ala Ala Ser Asp Ile Pro Asn

145 150 155 160

Gln Asp Gly His Leu Lys Phe Val Tyr His Lys Asp Gly Thr Gly Thr

165 170 175

His Ala Phe Arg Phe Ala Lys Asn Asn Glu Asp Ala Glu Asn Pro Leu

180 185 190

Arg Thr Phe Val Phe Pro Thr Val Val Ser Trp Tyr Thr Met Lys Gly

195 200 205

Asp Gly Ile Asp Asn Ala Gly Leu Arg Asp Lys Met Asn Ser His Asn

210 215 220

Tyr Gly Ser Ala Thr Met Pro Ile Lys Asp Ser Thr Phe Leu Asn Asn

225 230 235 240

Ile Asn Asn Phe Arg Pro Ser Gly Phe Pro Val Phe Thr Gln Asp Ser

245 250 255

Val Asn Asn Ser Gln

260

<210> 14

<211> 262

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 14

Met Asn Trp Ser Ala Met Leu Ser Leu Cys Val Ala Thr Ala Leu Ser

1 5 10 15

Ser Val His Ala Asp Asp Phe Pro Lys Leu Asp Glu Ala Met Pro Arg

20 25 30

Asn Ile Asp Ile Arg Ser Ile Thr Pro Met Ile Asp Thr Asp Thr Asp

35 40 45

Ser Cys Leu Pro Ser Ala Met Val Ser Arg Thr Gly Ala Gln Asn Ala

50 55 60

Gly Leu Asp Pro Glu Trp Ala Ser Leu Val Lys Gly Cys Arg Asn Asn

65 70 75 80

Asp Phe Met Asn Leu Ser Asn Thr Tyr His Arg Tyr Val Cys Gln Asn

85 90 95

Asn Asn Gly Asn Gln Tyr Cys Ala His Leu Phe Ala Tyr Tyr Phe Leu

100 105 110

Lys Asp Gln Ser Val Asn Leu Ile Pro Ser Gly His Lys Asn Asp Leu

115 120 125

Glu Gln Val Ala Val Trp Thr Ile Asn Gly Arg Val Ser His Gly Ser

130 135 140

Tyr Ser Ala His Gly Lys Met Tyr Asn Lys Ala Ala Ser Glu Ile Pro

145 150 155 160

Thr Gln Asp Gly His Leu Lys Phe Val Tyr His Lys Asp Gly Gly Ser

165 170 175

Thr His Cys Phe Arg Phe Ala Gly Asn Asn Glu Phe Ala Glu Asn Pro

180 185 190

Ala Gly Thr Phe Val Thr Pro Thr Val Val Ser Trp Tyr Thr Met Lys

195 200 205

Gly Asp Gly Ile Asp Asn Gly Gly Leu Arg Asp Arg Met Asn Asn Tyr

210 215 220

Asn Tyr Gly Ser Ala Thr Met Pro Ile Lys Asp Ser Thr Phe Leu Asn

225 230 235 240

Asn Val Asn Thr Tyr Lys Pro Gly Glu Phe Pro Val Phe Thr Gln Asp

245 250 255

Ser Val Asn Asn Ser Gln

260

<210> 15

<211> 259

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 15

Met Lys Trp Ile Ala Leu Leu Ala Ser Ile Ala Leu Leu Gly Arg Val

1 5 10 15

Arg Gly Asp Asp Phe Pro Lys Leu Asp Ala Ala Pro Pro Lys Asn Ile

20 25 30

Asp Ile Phe Ser Ile Thr Pro Val Leu Asp Met Asp Thr Asp Ser Cys

35 40 45

Leu Pro Ser Ala Ala Ile Ser Arg Thr Gly Ala Gln Asn Gly Gly Leu

50 55 60

Asn Pro Ser Gly Ser Ile Thr Gly Gly Cys Arg Arg Ser Asp Phe Met

65 70 75 80

Asn Tyr Ser Asn Thr Tyr His Arg Tyr Ala Cys Ile Ser Ser Asn Gly

85 90 95

Asn Gln Tyr Cys Gly His Phe Phe Ala Tyr Tyr Phe Leu Lys Asp Gln

100 105 110

Val Leu Ser Thr Val Gly Gly Gly His Arg His Asp Ile Glu Gln Val

115 120 125

Gly Ile Trp Thr Thr Asn Gly Arg Val Thr His Gly Gly Phe Ser Ala

130 135 140

His Gly Ala Met Thr Asn Leu Pro Ala Asn Gln Ile Pro Gln Glu Gly

145 150 155 160

Gly Arg Leu Lys Phe Val Tyr His Lys Asp Gly Val Ala Thr His Ala

165 170 175

Ile Arg Phe Ala Lys Thr Asn Glu Ala Val Glu Asn Pro Thr Gly Ala

180 185 190

Phe Val Val Pro Thr Leu Val Ser Trp Phe Thr Met Lys Ala Asp Ser

195 200 205

Ile Asn Asn Ala Gly Leu Arg Asn Asn Leu Asn Ser Phe Asp Tyr Gly

210 215 220

Asn Ala Asn Met Pro Ile Lys Asp Ser Asn Phe Leu Asn Ser Leu Asn

225 230 235 240

Lys Phe Arg Pro Ala Glu Tyr Pro Ala Phe Thr Gln Ala Ser Val Asp

245 250 255

Asn Ser Gln

<210> 16

<211> 262

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 16

Met Asn Trp Ser Ala Ile Leu Ser Leu Cys Val Ala Ala Ala Leu Gly

1 5 10 15

Ser Val His Ala Asp Asp Phe Pro Lys Leu Asp Glu Ala Met Pro Arg

20 25 30

Asn Ile Asp Ile Arg Ala Ile Thr Pro Val Val Asp Thr Asp Thr Asp

35 40 45

Gly Cys Leu Pro Ser Ala Met Val Ser Arg Thr Gly Gln Gln Asn Gly

50 55 60

Gly Leu Ala Pro Gly Trp Ser Asn Leu Thr Lys Gly Cys Arg Asn Asn

65 70 75 80

Asp Phe Met Asn Leu Ser Asn Thr Tyr His Arg Tyr Val Cys Gln Asn

85 90 95

Asn Asn Gly Asn Gln Tyr Cys Ala His Leu Phe Ala Tyr Tyr Phe Leu

100 105 110

Lys Asp Gln Ser Val Asn Leu Ile Pro Ser Gly His Lys His Asp Leu

115 120 125

Glu Gln Val Ala Val Trp Thr Ile Asn Gly Arg Val Ser His Gly Ser

130 135 140

Tyr Ser Ala His Gly Lys Met Tyr Ser Lys Pro Ala Ser Glu Ile Pro

145 150 155 160

Thr Gln Asp Gly His Leu Lys Phe Val Tyr His Lys Asp Gly Gly Ser

165 170 175

Thr His Cys Phe Arg Phe Ala Gly Asn Asn Glu Phe Ala Glu Asn Pro

180 185 190

Ala Gly Thr Phe Val Thr Pro Thr Leu Val Ser Trp Tyr Thr Met Lys

195 200 205

Gly Asp Gly Ile Asp Asn Gly Gly Leu Arg Asp Lys Met Asn Asn Tyr

210 215 220

Asn Tyr Gly Ser Ala Ser Met Pro Ile Lys Asp Gly Ser Phe Leu Asn

225 230 235 240

Asn Val Asn Thr Tyr Lys Pro Gly Glu Phe Pro Thr Phe Thr Gln Asp

245 250 255

Ser Val Asn Asn Ser Gln

260

<210> 17

<211> 262

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 17

Met Lys Trp Ser Ala Ile Leu Ser Leu Cys Val Ala Ala Val Leu Gly

1 5 10 15

Ser Ala His Ala Asp Asp Phe Pro Lys Leu Asp Glu Ala Met Pro Lys

20 25 30

Asn Val Asp Ile Arg Gly Ile Thr Pro Val Ile Asp Thr Asp Met Asp

35 40 45

Ser Cys Leu Pro Ser Ala Met Val Ser Arg Thr Gly Gln Gln Asn Gly

50 55 60

Gly Leu Glu Pro Ala Trp Ala Ser Leu Val Lys Gly Cys Arg Asn Asn

65 70 75 80

Asp Phe Met Lys Leu Ser Asn Thr Tyr His Arg Tyr Val Cys Gln Thr

85 90 95

Ser Asn Asn Asn Gln Tyr Cys Ala His Leu Phe Ala Tyr Tyr Phe Leu

100 105 110

Lys Asp Gln Ser Val Asn Leu Ile Pro Ser Gly His Lys His Asp Leu

115 120 125

Glu Tyr Val Ala Val Trp Thr Ile Asn Gly Arg Met Thr His Gly Ser

130 135 140

Tyr Ser Ala His Gly Lys Thr Tyr Asn Lys Pro Ala Thr Glu Ile Pro

145 150 155 160

Thr Gln Asp Gly His Leu Lys Phe Val Tyr His Lys Asp Gly Gly Leu

165 170 175

Thr His Cys Phe Arg Phe Ala Gln Lys Asn Glu Asn Ala Glu Asn Pro

180 185 190

Ala Gly Thr Phe Val Leu Pro Thr Leu Ala Ser Trp Tyr Thr Met Lys

195 200 205

Gly Asp Gly Ile Asp Asn Ala Gly Leu Arg Asn Lys Leu Asn Ser Tyr

210 215 220

Ser Tyr Gly Asn Ala Asn Met Pro Leu Lys Asp Gly Lys Phe Leu Asp

225 230 235 240

Lys Val Asn Ser Phe Lys Pro Ser Glu Phe Pro Ser Phe Thr Glu Asp

245 250 255

His Ala Lys Asn Ser Gln

260

<210> 18

<211> 260

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 18

Met Lys Trp Ala Ala Leu Leu Val Phe Ser Ala Ala Leu Leu Ser Thr

1 5 10 15

Val Arg Gly Asp Asp Phe Pro Lys Leu Asp Ala Ala Pro Pro Lys Asn

20 25 30

Ile Asp Ile Ser Ser Ile Thr Pro Val Leu Asp Met Asp Thr Asp Ser

35 40 45

Cys Leu Pro Ser Ala Ala Ile Ser Arg Ser Gly Gln Gln Asn Gly Gly

50 55 60

Leu Lys Pro Thr Gly Lys Ile Thr Gly Gly Cys Arg Arg Gly Asp Phe

65 70 75 80

Met Asp Phe Ser Asn Thr Tyr His Arg Tyr Ala Cys Val Asn Gly Asn

85 90 95

Gly Asn Gln Tyr Cys Gly His Phe Phe Ala Tyr Tyr Phe Leu Lys Asp

100 105 110

Gln Ala Val Pro Thr Phe Gly Gly Gly His Arg His Asp Leu Glu Gln

115 120 125

Ile Gly Ile Trp Thr Val Asn Gly Arg Val Thr His Gly Gly Phe Ser

130 135 140

Ala His Gly Lys Met Glu Asn Arg Pro Val Asp Gln Ile Pro Gln Glu

145 150 155 160

Gly Gly Arg Leu Lys Phe Val Tyr His Lys Asp Gly Val Val Thr His

165 170 175

Ala Ile Arg Phe Gly Lys Thr Asn Glu Gly Ala Glu Asn Pro Thr Gly

180 185 190

Ala Phe Val Val Pro Thr Leu Val Ser Trp Phe Thr Met Lys Gly Asp

195 200 205

Thr Ile Asp Asn Ala Gly Leu Arg Asn Asn Leu Asn Ser Phe Asp Tyr

210 215 220

Gly Asn Ala Asn Met Pro Ile Lys Asp Asn Asn Phe Leu Ser Asn Leu

225 230 235 240

Asn Lys Phe Arg Pro Ser Asp Phe Pro Glu Phe Thr Gln Glu Ser Ile

245 250 255

Asp Thr Ser Gln

260

<210> 19

<211> 265

<212> PRT

<213> 寡雄腐霉(Pythium oligandrum)

<400> 19

Met Lys Pro Ala Thr Phe Leu Thr Phe Leu Ala Ile Gly Leu Gln Ala

1 5 10 15

Thr Ala Thr Tyr Ala His Asp Phe Pro Lys Met Asp Glu Ala Leu Pro

20 25 30

Lys Asn Ile Asp Ile Arg Ser Ile Thr Pro Leu Leu Asp Phe Asp Glu

35 40 45

Asp Ser Cys Leu Pro Ser Thr Ala Val Ser Arg Glu Gly Lys Gln Asn

50 55 60

Gly Gly Leu Lys Thr Thr Gly Asn Ile Ala Gly Gly Cys Arg Asp Lys

65 70 75 80

Glu Asp Phe Leu Glu Tyr Ser Asn Thr Tyr Tyr Arg Tyr Val Cys Ala

85 90 95

Asn Tyr Glu Gly Lys Lys Tyr Cys Gly His Met Phe Ala Leu Tyr Phe

100 105 110

Gln Lys Asp Gln Ala Thr Leu Leu Gly Leu Gly His Arg His Asp Trp

115 120 125

Glu His Val Thr Val Trp Thr Thr Asp Gly Ile Val Thr His Gly Ser

130 135 140

Ala Ser Ala His Gly Gly Gly Lys Thr Ala Ser Phe Gly Ala Ile Pro

145 150 155 160

Asn Glu Asp Gln His Leu Leu Phe Val Tyr His Lys Asp Asp Val Gln

165 170 175

Thr His Ala Met Arg Phe Ala Asp Ala Asp Asp Gln Ala Glu Ala Glu

180 185 190

Asn Pro Tyr Lys Gln Phe Val Leu Pro Pro Val Val Ser Trp Tyr Ser

195 200 205

Met Gln Gly Asp Gly Ala Leu Asp Asn Glu Thr Leu Arg Asn Met Met

210 215 220

Asn Gly Leu Asn Phe Gly Ser Ala His Cys Pro Leu Leu Asp Gly Asp

225 230 235 240

Phe Leu Arg Asn Leu Asn Ser Leu Lys Pro Glu Gly Tyr Pro Thr Phe

245 250 255

Thr Asp Ala Asp Met Ile Gln Ser Lys

260 265

<210> 20

<211> 266

<212> PRT

<213> 缠器腐霉(Pythium periplocum)

<400> 20

Met Lys Phe Ala Ser Ile Leu Ala Phe Leu Thr Val Gly Ile Gln Val

1 5 10 15

Ser Ile Thr Gln Ala His Asp Phe Pro Arg Leu Asp Glu Ala Met Pro

20 25 30

Lys Gly Ile Asp Ile Lys Ala Ile Leu Pro Leu Leu Asp Phe Asp Asp

35 40 45

Asp Ser Cys Tyr Pro Ser Ala Ala Val Ser Arg Val Gly Arg Gln Asn

50 55 60

Gly Gly Leu Arg Thr Thr Gly Ser Ile Thr Gly Gly Cys Arg Asn Lys

65 70 75 80

Glu Phe Leu Asp Tyr Ser Asn Ser Tyr His Arg Tyr Ala Cys Arg Asp

85 90 95

His Glu Lys Lys Lys Tyr Cys Gly His Met Phe Ala Leu Tyr Phe Gln

100 105 110

Lys Asp Gln Met Val Asp Gly Leu Asp Pro Phe Gly His Arg His Asp

115 120 125

Phe Glu Tyr Val Ala Val Trp Thr Thr Asp Gly Val Val Thr His Gly

130 135 140

Ser Ala Ser Ala His Gly Asp Ala Ala Val Lys Pro Ile Asp Gln Ile

145 150 155 160

Pro His Ser Gly Glu His Ile Lys Phe Val Phe His Lys Asp Glu Pro

165 170 175

Thr Thr His Ser Met Arg Phe Ala Glu Lys Asn His Asp Glu Asp Ala

180 185 190

Glu Asn Ser Tyr Gly Asp Phe Val Leu Pro Ala Val Val Ser Trp Tyr

195 200 205

Thr Met Gln Gly Asp Gly Asn Met Thr Asn Asp Ala Leu Arg Gly Leu

210 215 220

Met Asn Ser Leu Asp Phe Gly Asp Ala His Asn Pro Leu Leu Asp Gly

225 230 235 240

Asp Phe Leu Asp Asn Leu Asn Glu Ala Lys Pro Lys Glu Tyr Pro Gln

245 250 255

Phe Asp Asp Glu Asp Val Lys Gly Ser Gln

260 265

<210> 21

<211> 265

<212> PRT

<213> 缠器腐霉(Pythium periplocum)

<400> 21

Met Lys Pro Ala Thr Phe Leu Thr Phe Leu Ala Ile Gly Leu Gln Ala

1 5 10 15

Thr Ala Thr Tyr Ala His Asp Phe Pro Lys Met Asp Glu Ala Leu Pro

20 25 30

Lys Asn Ile Asp Ile Arg Ser Ile Thr Pro Leu Leu Asp Phe Asp Glu

35 40 45

Asp Ser Cys Leu Pro Ser Thr Ala Val Ser Arg Glu Gly Lys Gln Asn

50 55 60

Gly Gly Leu Lys Thr Ser Gly Asn Ile Ala Gly Gly Cys Arg Asp Lys

65 70 75 80

Glu Asp Phe Leu Glu Tyr Ser Asn Thr Tyr His Arg Tyr Val Cys Ala

85 90 95

Asn Tyr Glu Gly Lys Lys Tyr Cys Gly His Met Phe Ala Leu Tyr Phe

100 105 110

Gln Lys Asp Gln Ala Thr Leu Leu Gly Leu Gly His Arg His Asp Trp

115 120 125

Glu His Val Thr Val Trp Thr Thr Asp Gly Ile Val Thr His Gly Ser

130 135 140

Ala Ser Ala His Gly Gly Gly Lys Thr Ala Ser Phe Asp Ala Ile Pro

145 150 155 160

Asn Glu Asp Gln His Leu Leu Phe Val Tyr His Lys Asp Asp Val Gln

165 170 175

Thr His Ala Met Arg Phe Ala Asp Ala Asp Asp Gln Ala Glu Val Glu

180 185 190

Asn Pro Tyr Leu Gln Phe Val Leu Pro Pro Val Val Ser Trp Tyr Ser

195 200 205

Met Gln Gly Asp Gly Ala Leu Asp Asn Glu Thr Leu Arg Asn Met Met

210 215 220

Asn Arg Leu Asn Phe Gly Ser Ala His Cys Pro Leu Leu Asp Gly Asp

225 230 235 240

Phe Leu Arg Asn Leu Asn Ser Leu Lys Pro Glu Gly Tyr Pro Thr Phe

245 250 255

Thr Asp Ala Asp Met Ile Gln Ser Lys

260 265

<210> 22

<211> 261

<212> PRT

<213> 缠器腐霉(Pythium periplocum)

<400> 22

Met Lys Leu Leu Thr Ile Leu Ser Val Val Cys Ala Ala Val Ala Gly

1 5 10 15

Ser Val Arg Ala Asp Asp Phe Pro Lys Leu Asp Glu Ala Trp Pro Lys

20 25 30

Asn Val Asp Ile Arg Ser Ile Thr Pro Val Phe Asp Met Asp Thr Asp

35 40 45

Ser Cys Leu Pro Ser Ala Met Val Ser Arg Thr Gly Gln Gln Asn Gly

50 55 60

Gly Leu Lys Pro Ser Gly Gln Ile Val Gly Lys Cys Tyr Asn His Asp

65 70 75 80

Phe Met Asn Leu Ser Asn Ser Tyr His Arg Tyr Ala Cys Gln Asn Ser

85 90 95

Asn Gly Ser Gln Tyr Cys Ala His Leu Phe Ala Tyr Tyr Phe Leu Lys

100 105 110

Asp Gln Lys Ile Asn Phe Ile Glu Ser Gly His Arg His Asp Leu Glu

115 120 125

Gln Leu Ala Val Trp Thr Ile Asn Gly Arg Val Ser His Gly Ser Tyr

130 135 140

Ser Ala His Gly Lys Met Tyr Ala Lys Ala Ala Ser Asp Ile Pro Asn

145 150 155 160

Gln Asp Gly His Leu Lys Phe Val Tyr His Lys Asp Gly Ala Gly Thr

165 170 175

His Ala Phe Arg Phe Ala Lys Asn Asn Glu Asp Ala Glu Asn Pro Leu

180 185 190

Arg Thr Phe Val Phe Pro Thr Val Val Ser Trp Tyr Thr Met Lys Gly

195 200 205

Asp Gly Ile Asp Asn Ala Gly Leu Arg Asp Lys Met Asn Ser Tyr Asn

210 215 220

Tyr Gly Ser Ala Thr Met Pro Ile Lys Asp Ser Thr Phe Leu Asn Asn

225 230 235 240

Ile Asn Asn Phe Arg Pro Ser Gly Phe Pro Val Phe Thr Gln Asp Ser

245 250 255

Val Asn Asn Ser Gln

260

Claims (9)

1.一种生防腐霉菌中诱导坏死和活性氧积累的NLP类基因，其特征在于，该NLP类基因为PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8基因中的至少一种，所述PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8基因的核苷酸序列依次分别如SEQ ID NO.1～11所示。

2.由权利要求1所述的NLP类基因编码的蛋白，其特征在于，所述PyolNLP1、PyolNLP2、PyolNLP3、PyolNLP4、PyolNLP5、PyolNLP6、PyolNLP7、PyolNLP9、PypeNLP4、PypeNLP5和PypeNLP8基因编码的蛋白的氨基酸序列依次分别如SEQ ID NO.12—22所示。

3.含有权利要求1所述NLP类基因的表达盒、重组表达载体、转基因细胞系或转基因重组菌。

4.根据权利要求3所述的重组表达载体，其特征在于，该重组表达载体的出发载体为表达载体PBIN-PLUS。

5.根据权利要求4所述的重组表达载体，其特征在于，该重组表达载体为将权利要求1所述的NLP类基因插入PBIN-PLUS载体的SmaI酶切位点间所得。

6.权利要求1中所述的NLP类基因或权利要求2中所述的蛋白在诱导植物产生坏死和活性氧积累中的应用。

7.权利要求4或5所述的重组表达载体在诱导植物产生坏死和活性氧积累中的应用。

8.权利要求1中所述的NLP类基因或权利要求2中所述的蛋白在开发植物免疫诱抗剂中的应用。

9.权利要求4或5所述的重组表达载体在开发植物免疫诱抗剂中的应用。

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