CN104561024A - Rapid identification of MLO (Mycoplasma Like Organism) powdery mildew resistant genes of melon - Google Patents
Rapid identification of MLO (Mycoplasma Like Organism) powdery mildew resistant genes of melon Download PDFInfo
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- CN104561024A CN104561024A CN201310517585.XA CN201310517585A CN104561024A CN 104561024 A CN104561024 A CN 104561024A CN 201310517585 A CN201310517585 A CN 201310517585A CN 104561024 A CN104561024 A CN 104561024A
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Abstract
The invention relates to rapid identification of MLO (Mycoplasma Like Organism) powdery mildew resistant genes of a melon, relates to comparative genomics of plants, genetics, bioinformatics and other disciplinary knowledge, and belongs to the field of biotechnology science of plants. The rapid identification is mainly carried by the following steps: 1) downloading the whole genome sequence of the melon, and acquiring MLO genes; 2) identifying the MLO genes; 3) obtaining the phylogenetic relationship of MLO genes; 4) comparing MLO powdery mildew resistant genes. By virtue of adopting the rapid identification, the powdery mildew resistant genes mining cycle of the melon can be effectively reduced, which is beneficial to the rapid identification of powdery mildew resistant genes; the identified candidate powdery mildew resistant genes are utilized to develop corresponding coseparation functional markers (such as SNR and SCAR) and can also be quickly used for assistant selection of molecular markers resistant to powdery mildew resistant genes, and the accuracy is high; in combination with other anti-disease gene molecular markers, a plurality of resistive breeding materials can be created, the breeding periods can be reduced, and the breeding efficiency can be increased; the foundation is provided for describing a powdery mildew resistant molecular mechanisms of the melon.
Description
Technical field
The present invention is by means of muskmelon order-checking whole genome sequence, utilize the method Rapid identification melon powdery mildew genes such as plant comparative genomics, genetics, information biology and candidate gene strategy, be mainly concerned with the download of muskmelon whole genome sequence, the qualification of candidate gene, the comparison of gene, the means such as cluster, and then identify powdery mildew gene, belong to Plant Biotechnology scientific domain.
Background technology
Muskmelon is also known as sweet melon or muskmelon.Muskmelon is gained the name because taste is sweet, attacks people therefore have another name called muskmelon due to delicate fragrance.Muskmelon is melon and fruit of relieving summer heat in the summertime, and its nutritive value can match in excellence or beauty with watermelon.According to surveying and determination, muskmelon is except the content of moisture and protein is lower than except watermelon, and other nutritive ingredients are all no less than watermelon, and aromatoising substance, mineral substance, sugar and ascorbic content are then apparently higher than watermelon.Eat muskmelon more, be conducive to the activity of human heart and liver and intestinal tract, promote internal secretion and hematopoietic function.Motherland's medical science confirms the remarkable efficacy that muskmelon has " heat of relieving summer heat separates polydipsia, diuresis ".The production of Powdery Mildew serious harm muskmelon, all can be injured, especially be injured the heaviest with Later growth from seedling stage to harvesting time.Mainly to cause harm blade.Just existing white mildew on blade face, after gradually to surrounding expansion, form subcircular patch, when condition is suitable for, mildew expansion is very rapid, and scab is fused into a slice mutually, causes whole blade covered with the white meal of one deck, serious melon face also can grow white powder thing.At present aborning, the control of melon powdery mildew is mainly by spraying fungicide, but life-time service medicine can cause environmental pollution and Biological Strains of The Pest variation, and therefore promoting disease-resistant variety is method the most safely and effectively.
MLO type disease-resistant gene is the disease-resistant gene that in plant, a class is special.Investigator finds that MLO gene pairs powder mildew resistance starts from 1937-1938 the earliest, fritz acquires the barley of a lot of kind in Ethiopia, two strains wherein all have efficient resistance to all known physiological strains of Powdery Mildew (Blumeria.graminisf.sp.hexlei, Bgh).Further research shows, in barley, the recessive mutation mlo of MLO gene can make barley produce resistance that is lasting, wide spectrum to the physiological strain of nearly all known barley powdery mildew bacteria.Therefore the MLO type disease-resistant gene excavated in plant has great importance to Biology of Plant-Powdery Mildew Interaction breeding.At present, the method for clone plant disease-resistant gene mainly contains transposon tagging, map based cloning method etc.Therefore, how the MLO type disease-resistant gene of Rapid identification muskmelon will become the important prerequisite of powdery mildew of melon breeding.
Plant comparative genomics (Comparative Genomics) is the subject that the information obtained based on Genome Atlas and order-checking infers other biological genomic number gene, position, function, expression mechanism and spore.Between Land use models Plant Genome and other Plant Genome on coded sequence and structural homology, clone's other plant gene, disclose gene function and molecular mechanism, illustrate spore relation and genomic immanent structure, the control of the Powdery Mildew to cocoa tree can be advanced faster.The importance of plant comparative genomics is that it links together different subjects, different biological species, has erected the bridge between fundamental research and applied research.Across kind, across belonging to, genome comparison transboundary understands the relation of gene and genomic structure, gene structure and function and DNA to us and changes and how to cause species diversity etc. to be of great significance.
The method that this patent adopts and thinking: the research of model plant arabidopsis gene group has disclosed the function of MLO type gene, utilize the homology clone muskmelon MLO type disease-resistant gene in its order of gene, according to the feature of the superiority in model plant Arabidopis thaliana experimental system and known MLO type disease-resistant gene, " seizure " powdery mildew of melon gene fast.This patent describes premised on muskmelon whole genome sequence, in conjunction with knowledge such as comparative genomics, genetics, genomics, information biology and candidate gene strategy, excavates powdery mildew gene fast.
Summary of the invention
Technical problem
The object of this invention is to provide a kind of by conjunction with knowledge such as plant comparative genomics, plant genetics, genomics and information biology, excavate powdery mildew of melon gene fast.Its result can be used for the exploitation of melon powdery mildew gene compact linkage molecule mark on the one hand, carries out molecular marker assisted selection breeding, on the other hand also for other crop powdery mildew gene identification provide reference frame.
Technical scheme
Cardinal principle: first Biology of Plant-Powdery Mildew Interaction gene (MLO) is cloned from barley, research finds that this gene is the special disease-resistant gene of a class, is different from most of NBS (nucleotide-binding site) the type disease-resistant gene of previously clone; Subsequently, investigator has cloned powdery mildew gene in succession from the plants such as tomato, Arabidopis thaliana, pea, capsicum, Root or stem of Littleleaf Indianmulberry, and what research found these genes encodings is all MLO type disease-resistant gene.Subsequently, by test of many times, numerous investigator confirms that MLO type disease-resistant gene has become the distinctive class mildew-resistance gene of plant.Further discovery, plant MLO type gene is a gene family; And Phylogenetic Relationships analysis discovery is carried out to the MLO gene family deriving from different plant species, in different plant species mildew-resistance gene always cluster together, become a class, this class MLO gene all has the characteristic feature of mildew-resistance gene sequence.Completing of muskmelon gene order-checking provides a convenient approach for excavating powdery mildew gene.Therefore, can by means of the Phylogenetic Relationships of MLO gene family in the muskmelon full-length genome checked order and the MLO powdery mildew gene of having cloned and for maintaining the conservation of amino acids of powdery mildew gene MLO critical function to identify melon powdery mildew gene.
Key step is as follows:
1) download of muskmelon whole genome sequence and the collection of MLO type gene thereof
First muskmelon whole genome sequence is downloaded from muskmelon sequenced genes group database (http://melonomics.net/); Use " DNATOOLS " software to the muskmelon full-length genome amino acid sequence data building database obtained, then pfam database (protein family database is used, http://pfam.janelia.org/search/sequence) in hidden Markov model (HMM) Blastp (E-value=0.001) sequence alignment is carried out to the aminoacid sequence of MLO structural domain and the muskmelon full-length genome amino acid sequence database to have set up, preliminary screening goes out candidate gene sequence.Secondly, utilize the MLO type gene order announced, BLAST comparison is carried out to muskmelon genome database, obtain candidate gene sequence.
2) qualification of muskmelon MLO type gene family
By the candidate gene of homologous amino acid sequence obtained in the above results, analyzed by Pfam (E-value=1.0), remove the gene order (Fig. 1) without ' MLO ' structural domain.The ClustalW instrument (Multiple Sequence Alignment program) Candidate Disease Resistant Genes sequence provided by MEGA3.1 software again carries out Multiple Sequence Alignment, removes tumor-necrosis factor glycoproteins.
3) by the muskmelon MLO type powdery mildew gene of the Phylogenetic Relationships qualification candidate of plant MLO type gene
Because previous research is verified, dicotyledons MLO type powdery mildew gene is positioned at plant MLO Phylogenetic Tree same district group, therefore in Phylogenetic Relationships research, we are MLO type mildew-resistance gene with muskmelon MLO type gene together with the cluster analysis of the MLO type gene family of Arabidopis thaliana with some other crops, to obtain the powdery mildew of melon gene (Fig. 2) of candidate.
4) comparison of melon powdery mildew gene and known plant MLO powdery mildew gene
BioXM2.6 software is utilized to convert the aminoacid sequence of the MLO powdery mildew gene of the MLO type powdery mildew gene of muskmelon candidate and Arabidopis thaliana, tomato, pea, barley the file of Fasta form to, these files are imported BioEdit7.0 software, use Clustal software in this software to carry out Multiple Sequence Alignment, disclose the conservative property in candidate's powdery mildew gene important amino acid residue and region.Thus identify the powdery mildew gene (Fig. 3) of muskmelon candidate further.
Positively effect of the present invention:
1) shorten the melon powdery mildew gene excavating cycle, be conducive to the Rapid identification of powdery mildew gene.Mildew-resistance gene not only takes time and effort, efficiency is low to adopt ordinary method (map based cloning, transposon tagging etc.) to excavate, and is difficult to successfully.The present invention is based on plant comparative genomics, genetics, bioinformatics method and excavate melon powdery mildew gene fast, not only can shorten the time, powdery mildew gene determination rates can also be improved.
2) muskmelon (Cucumis melo) is one of important vegetables in the whole world.Because muskmelon hereditary basis is narrow, Germplasm Resources Diversity is low, therefore by conventional molecule marker (RAPD, ISSR, SSR, AFLP etc.) qualification melon powdery mildew genetic comparison difficulty.The candidate's powdery mildew gene exploitation be tested and appraised is divided into from functional indicia (SNP, SCAR etc.) accordingly, can fast for the molecular marker assisted selection of disease-resistant gene, and accuracy is high.
3) initiative of multiresistance breeding material.Based on the functional molecular marker of the powdery mildew gene exploitation of new qualification, ins conjunction with the molecule marker of other disease-resistant genes after positioning, carry out the initiative of multiresistance breeding material, can shortening the breeding cycle, raising breeding efficiency.
4) lay a good foundation for setting forth powdery mildew of melon molecular mechanism.The qualification of powdery mildew of melon gene, by transgenic technology, RNAi, virus induced gene silencing (virus induced gene silencing, VIGS) molecular mechanism of the Effect of Anti Powdery Mildew such as technology provides genetic resources, is conducive to the mechanism of action setting forth powdery mildew of melon fast.
Accompanying drawing explanation
The qualification of Fig. 1 muskmelon MLO gene;
What this figure showed is 14 MLO type gene identification results, and each gene is containing ' MLO ' conserved domain.
The Phylogenetic Relationships analysis of Fig. 2 plant MLO gene family and the qualification of muskmelon MLO type powdery mildew gene thereof;
Arabidopis thaliana is the model plant of plant science research, in phylogenetic tree construction, the powdery mildew gene (PsMLO) of 15 MLO type genes (wherein 3 genes are powdery mildew genes: AtMLO02, AtMLO06 and AtMLO12) of Arabidopis thaliana, Gene against Powdery Mildew in Tomato (S1MLO), barley powdery mildew gene (HvMLO) and pea is selected to and the analysis of muskmelon MLO type gene clusters.Identify the MLO type powdery mildew gene of 3 muskmelon candidates altogether.In figure, the gene of italic mark is exactly candidate's melon powdery mildew gene.
The compare of analysis of Fig. 3 muskmelon MLO type powdery mildew gene;
3 melon powdery mildew genes and barley (HvMLO), tomato (S1MLO), pea (PsMLO), Arabidopis thaliana powdery mildew gene (AtMLO02, AtMLO06 and AtMLO12) to compare, qualification powdery mildew infects the conservative type in amino-acid residue and the region played an important role.In figure, TM1-TM7 represents 7 revolving die regions of muskmelon MLO type powdery mildew gene; Black round dot represents that powdery mildew infects important amino-acid residue; CaMBD represents calmodulin CaM binding region; I and II represents and infects important amino acid region to powdery mildew.
Embodiment
The qualification of disease-resistant gene has important effect in the research of crop disease-resistant theory of heredity and disease-resistant variety seed selection.Present method Rapid identification can go out melon powdery mildew gene.Specific implementation process is as follows:
1) collection of muskmelon MLO type gene and qualification
In order to obtain the whole MLO type gene family member of muskmelon, we are first with the MLO type gene of Arabidopis thaliana, the mildew-resistance MLO gene order of tomato, pea, capsicum, rose, capsicum, Root or stem of Littleleaf Indianmulberry builds HMM model, receives rope MLO type gene from muskmelon genome sequence, secondly the MLO gene order delivered in Different Crop as target sequence (from DFCI database: TC171015, TC267529, DFCI:TC327983, TC289653, TC312087, TC132500, TC133436, TC317623, TC317025, TC315947, TC325903, TC315944, TC315912, TC322759, TC322059, TC330654, TC282713, TC293173, TC281861, TC283253, TC283383, TC285032, TC290021, TC302716, TC283487, TC282866, TC283441, TC281428, TC285118, TC285090, from GenBank database: AY967408, AF384145, AF384144, AY029312-AY029315, AY029317-AY029319, Z95352, AF369563-AF369565, AF369567, AF369569-AF369576, Z83834, Z95496, AY581255), BLAST comparison is carried out to muskmelon database (http://www.phytozome.net/search.php), select the highest sequence of similarity to download, obtain the MLO type gene (MELO3C000169P1MELO3C005038P1 of 14 candidates altogether, MELO3C005044P1, MELO3C007979P1, MELO3C012438P1, MELO3C013709P1, MELO3C013761P1, MELO3C016709P2, MELO3C019435P1, MELO3C021515P2, MELO3C022486P1, MELO3C023219P1, MELO3C025761P1, MELO3C026525P1).
2) qualification of muskmelon MLO type gene family
In order to verify these MLO gene accuracys further, we have carried out the qualification of conserved domain " MLO " to the MLO gene of these 14 candidates.With the aminoacid sequence of the MLO type gene of each candidate for benchmark, PFAM (http://pfam.sanger.ac.uk/) website is carried out the qualification of ' MLO ' conserved domain, and concrete outcome is shown in Fig. 1.
3) the Phylogenetic Relationships analysis of muskmelon MLO type gene
In previous research, find that dicotyledons powdery mildew gene aggregates into district's group; Therefore, in phylogenetic tree construction, we have selected 15 MLO type genes (wherein 3 genes are powdery mildew genes: AtMLO02, AtMLO06 and AtMLO12) of model plant Arabidopis thaliana, the powdery mildew gene of Gene against Powdery Mildew in Tomato, barley powdery mildew gene and pea analyze with muskmelon MLO type gene clusters together with cluster analysis.Muskmelon MLO type gene and other crop powdery mildew gene protein sequences are carried out multisequencing connection to join (adopting Clustal X1.83 software to carry out), and utilize Genedoc software (http://www.nrbsc.org/gfx/genedoc/index.html) to show multisequencing to join the result of joining.Clustal multisequencing is joined the result of joining output in MEGA4.0 software, and utilize this software to construct adjacent tree (neighbor-joining, NJ) respectively, utilize Bootstrapping method to assess these evolutionary trees.Found that in dicotyledons mildew-resistance gene district group, there is the MLO type powdery mildew gene (see Fig. 2) of 3 muskmelon candidates.
4) comparison of muskmelon MLO type disease-resistant gene
In the research of barley MLO type powdery mildew gene, in succession found some important areas and single amino acids in research, they infect large wheat powdery mildew irreplaceable effect.In order to identify in the melon powdery mildew gene of 3 candidates, these important areas and amino acid whether high conservative, we have carried out compare of analysis to from 3 mildew-resistance genes (AtMLO02, AtMLO6 and AtMLO12) of Arabidopis thaliana, tomato powdery mildew gene (S1MLO), powdery mildew of pea gene (PsMLO).Find the powdery mildew gene of muskmelon 3 candidates and known MLO type powdery mildew gene 7 cross-film districts, 30 important amino acid, 1 calmodulin CaM binding region (CaMBD) and two important region (I and II) high conservatives (Fig. 3).
Claims (2)
1. powdery mildew of melon gene, it is characterized in that being selected from following 3 genes or its one of:
(1.)MELO3C005044P1
Amino acid:
Nucleotides:MAGAAGGKSLEQTPTWAVAVVCFVLLVVSIFIEYSLHLIGHWLKKRHKRALFEALEKIKSELMLLGFISLLLTVGQGPITEICIPQHVAATWHPCTKEREDEMNKEMEKSVQHLGHNRRRLLHLVGNGGSFRRSLAAAGGADKCAAKGKASFISAEGIHQLHIFIFVLAVFHVLYCVLTYALARAKMRSWKTWEKETKTAEYQFSHDPERFRFARDTSFGRRHLSFWTKNPAFMWIVCFFRQFVRSVPKVDYLTLRHGFIMAHLAPQSHTQFDFQKYINRSLEEDFKVVVGISPPIWFFAVLFLLSNTHGWRAYLWLPFIPLIILLLVGTKLQVIITKMALRIQERGEVVKGVPVVEPGDDLFWFNRPRLILYLINFVLFQNAFQVAFFAWTWYEFGLNSCFHEHIEDVVIRISMGVLVQILCSYVTLPLYALVTQMGSTMKPTIFNERVAEALRNWYHSARKHIKHNRGSVTPMSSRPATPTHSMSPVHLLRHYKSEVDSFHTSPRRFDTDRWDNDSPSPSRHVDGSSSSQPHVEMGGYEKDPVDSSSSQVDPIQPSRNRNQHEIHIGGPKDFSFDRVE
ATGGCGGGGGCAGCCGGTGGGAAGTCGCTCGAGCAAACACCGACATGGGCCGTTGCCGTTGTTTGCTTTGTTTTGCTCGTCGTCTCTATTTTCATCGAATATAGTCTCCATCTTATCGGACATTGGCTAAAGAAGAGACACAAACGGGCGTTATTTGAAGCATTAGAGAAGATCAAATCAGAGCTTATGTTATTGGGATTTATATCATTGCTACTGACGGTGGGGCAAGGACCAATAACGGAGATATGTATTCCACAACATGTAGCTGCAACCTGGCATCCATGTACAAAGGAAAGAGAAGATGAGATGAACAAAGAGATGGAGAAATCTGTGCAACATTTGGGTCATAATCGCCGGAGACTCCTTCATCTCGTCGGAAATGGTGGAAGTTTCCGGCGGAGTTTGGCCGCTGCGGGAGGAGCTGATAAATGTGCTGCCAAGGGTAAAGCTTCATTTATCTCAGCAGAAGGAATTCATCAACTTCATATCTTCATTTTTGTGTTGGCTGTTTTCCATGTTTTGTATTGTGTTCTAACTTATGCTTTGGCTAGAGCTAAGATGAGGAGTTGGAAAACATGGGAAAAAGAGACCAAAACTGCTGAATACCAATTCTCACATGATCCGGAGAGATTTAGGTTTGCAAGAGACACCTCATTTGGGAGAAGACATTTGAGCTTTTGGACCAAAAATCCTGCCTTCATGTGGATCGTTTGTTTCTTCAGACAATTTGTAAGATCTGTTCCAAAAGTTGATTACTTGACATTAAGACATGGGTTTATAATGGCACATTTAGCACCTCAAAGTCATACACAATTCGATTTTCAAAAATACATTAATAGATCCCTTGAAGAAGACTTCAAAGTTGTTGTGGGAATCAGCCCACCAATTTGGTTCTTTGCTGTTCTATTTCTCCTCTCAAACACTCACGGTTGGAGGGCATATCTATGGCTTCCATTCATCCCACTAATCATTTTGCTGTTGGTTGGAACAAAATTGCAAGTGATCATAACGAAAATGGCACTAAGAATACAAGAAAGAGGAGAAGTAGTGAAGGGCGTGCCGGTGGTGGAGCCTGGGGATGACCTTTTTTGGTTTAATCGACCTCGTCTTATTCTTTATCTCATCAACTTTGTTCTCTTTCAAAATGCCTTCCAAGTTGCTTTCTTTGCTTGGACTTGGTATGAGTTTGGGTTGAATTCTTGTTTCCACGAGCATATAGAGGATGTGGTGATCAGAATTTCCATGGGGGTGCTTGTACAAATCCTTTGCAGTTATGTTACTCTTCCTCTTTATGCACTAGTCACTCAGATGGGTTCAACAATGAAGCCAACTATATTCAACGAGAGAGTGGCAGAGGCCCTTCGCAATTGGTACCACTCGGCTCGAAAGCACATCAAACACAACCGCGGTTCGGTCACTCCAATGTCGAGCCGACCCGCCACCCCGACTCACAGCATGTCACCTGTCCACCTTCTCCGACACTACAAGAGTGAAGTCGATAGCTTCCACACCTCACCGAGAAGGTTCGACACCGACCGTTGGGACAATGATTCGCCCTCACCGTCTCGCCATGTTGATGGTTCGTCTTCCTCACAACCCCATGTCGAGATGGGAGGTTATGAAAAAGATCCCGTTGATTCAAGTTCGTCTCAAGTTGACCCGATTCAACCATCCCGAAACCGCAATCAACATGAGATTCATATTGGGGGCCCCAAAGACTTTTCATTTGATAGAGTTGAAT GA
(2.)MELO3C012438P1
Amino acid:
MAECGTEQRTLEDTSTWAVAVVCFFLVVISIFIEHVIHLTGKWLEKKHKPALVEALEKVKAELMLLGFISLLLTVGQDAVTQICVSKELAATWLPCAARAKAGVKVAKNSRLRLLEFLDPDYGSRRILASKGDDACAKRGQLAFVSAYGIHQLHIFIFVLAVFHVLYCIITLAFGRTKMSKWKAWEDETKTIEYQYYNDPARFRFARDTTFGRRHLSFWSRTPISLWIVCFFRQFFGSVTKVDYMTLRHGFIVAHLAPGSEVKFDFHKYISRSLEDDFKVVVGISPAMWLFAVLFILTNTNGWYSYLWLPFISLFIILLVGTKLHVIITHMGITIQERGHVVKGVPVVQPRDDLFWFGRPQLILFLIHFVLFMNAFQLAFFAWTTYAFTWRGCFHQRIEDIAIRLSMGVIIQVLCSYVTLPLYALVTQMGSNMRPTIFNDRVATALKNWHHSAKKNMKQHRNPDSTSPFSSRPTTPTHGMSPIHLLHKHQHGSTSPRLSDAEPDQWEELPPSSHHNRAHDNQDQQEQSETSREQEMTVQRPSSSETGSITRPARPHQEITRSPSDFSFAK
Nucleotide:
ATGGCTGAATGTGGAACAGAGCAGCGTACTTTGGAAGATACTTCAACTTGGGCTGTTGCGGTTGTTTGTTTTTTCTTGGTTGTTATTTCAATCTTCATTGAACATGTCATTCACCTCACTGGAAAGTGGCTGGAGAAAAAGCACAAGCCAGCTCTTGTTGAAGCTCTAGAAAAGGTTAAAGCAGAGCTTATGCTATTGGGATTTATATCCCTACTTCTAACGGTAGGCCAAGATGCTGTCACTCAAATTTGTGTTTCGAAGGAGCTTGCAGCAACTTGGCTTCCCTGTGCAGCAAGAGCTAAAGCAGGAGTAAAAGTTGCGAAGAACAGTCGTCTTAGACTTCTTGAATTTTTAGATCCTGACTATGGTTCGAGGCGTATTTTAGCCTCGAAAGGAGATGATGCATGCGCTAAGAGGGGCCAACTCGCTTTCGTGTCGGCATATGGAATCCATCAGCTCCATATTTTCATCTTCGTCTTGGCTGTCTTCCATGTCCTATATTGCATCATCACTTTGGCTTTCGGCAGAACAAAGATGAGCAAATGGAAGGCCTGGGAGGATGAAACCAAGACAATTGAATACCAGTACTATAATGATCCAGCAAGATTTAGATTTGCCAGAGATACTACGTTTGGACGCCGACACTTGAGCTTCTGGAGTCGTACACCAATTTCCCTCTGGATTGTTTGTTTCTTCAGACAATTCTTTGGATCAGTTACCAAGGTTGATTACATGACACTGAGACATGGATTCATTGTTGCACATCTAGCACCCGGAAGTGAAGTAAAATTTGATTTCCACAAATACATTAGCAGATCTCTGGAAGACGACTTTAAAGTTGTTGTGGGGATTAGTCCCGCAATGTGGCTATTTGCTGTTCTCTTCATCCTAACTAATACAAATGGGTGGTATTCATATCTATGGCTGCCTTTCATCTCCTTATTTATAATTCTATTGGTGGGAACAAAGCTCCATGTCATTATAACTCATATGGGATTGACAATTCAAGAAAGGGGTCATGTTGTGAAGGGTGTTCCGGTCGTTCAGCCTCGGGATGACCTGTTTTGGTTTGGCCGTCCACAACTTATTCTCTTCCTGATCCACTTTGTTCTCTTTATGAATGCATTTCAGCTTGCCTTCTTTGCTTGGACCACATATGCATTCACGTGGAGGGGTTGTTTCCATCAGCGAATTGAAGATATTGCCATCAGACTCTCAATGGGGGTTATCATACAAGTTCTCTGCAGTTATGTCACACTCCCACTCTATGCTTTAGTTACTCAGATGGGCTCTAACATGAGACCAACCATTTTCAACGACCGAGTGGCGACGGCATTGAAGAACTGGCACCACTCCGCCAAGAAGAACATGAAGCAGCATCGCAACCCAGACAGTACCTCACCATTCTCAAGCAGGCCAACAACTCCAACTCACGGCATGTCTCCTATTCACCTTCTGCACAAACATCAGCATGGCAGCACATCTCCCAGGCTATCCGATGCCGAACCCGATCAGTGGGAAGAGTTACCTCCTTCTTCACACCATAATAGAGCCCATGATAATCAAGATCAACAAGAACAATCTGAGACTAGTAGAGAACAGGAGATGACGGTTCAACGACCAAGTTCAAGTGAAACCGGTTCCATAACACGTCCTGCTCGCCCTCATCAGGAAATCACTAGGAGTCCATCGGACTTCTCATTTGCCAAATGA
(3.)MELO3C025761P1
Amino acid:
MAGGGAGRSLEETPTWAVAAVCFVLVLISIIIEHILHLIGKWLKKKHKRALYEALEKIKSELMLLGFISLLLTVGQSLITNVCIPPDVAATWHPCSPQREEELTKEADLVDSDHNRRKLLATVSHHVNATFRRSLAAAGGTDKCAAKGKVPFVSEGGIHQLHIFIFVLAVFHVLYCVLTLALGNAKMRSWKSWEKETRTVEYQFSHDPERFRFARDTSFGRRHLSFWTKSPFLIWIVCFFRQFVRSVPKVDYLTLRHGFVMAHLAPHSDQKFDFQKYIKRSLEEDFKVVVSISPPIWFFAVLFLLFNTNGWRAYLWLPFVPLIIVLLVGTKLQVIITKMALRIQERGEVVKGVPVVEPGDDLFWFNRPRLILYLINFVLFQNAFQLAFFAWTWKEFGMKSCFHEHTEDLVIRITMGVLVQILCSYVTLPLYALVTQV
Nucleotide:
ATGGCCGGAGGTGGCGCCGGAAGGTCCTTGGAAGAGACGCCGACATGGGCCGTCGCCGCCGTCTGCTTTGTTTTGGTTCTGATTTCTATTATCATCGAACACATTCTCCATCTCATCGGAAAGTGGCTAAAGAAGAAACACAAACGAGCTCTTTACGAAGCTCTAGAGAAGATCAAATCAGAACTGATGCTGTTGGGATTCATATCGCTGCTGCTGACGGTGGGACAAAGCCTAATCACAAATGTTTGTATACCACCAGACGTGGCAGCCACGTGGCATCCATGTAGTCCTCAAAGAGAAGAGGAATTAACTAAAGAAGCCGACCTGGTCGATTCCGACCACAATCGTCGGAAACTTCTCGCGACGGTCTCCCATCATGTCAACGCCACCTTCCGCCGTTCCCTCGCGGCTGCCGGTGGTACCGACAAATGTGCTGCCAAGGGTAAAGTTCCATTTGTATCGGAAGGGGGTATTCATCAGCTACATATATTCATCTTCGTATTGGCCGTTTTCCATGTTTTGTATTGTGTTCTAACTTTAGCTCTGGGCAATGCCAAGATGAGAAGTTGGAAGTCATGGGAGAAAGAGACTAGAACAGTGGAGTACCAATTCTCACACGATCCGGAACGGTTTCGATTTGCAAGAGACACGTCGTTTGGGAGAAGACATTTAAGCTTTTGGACAAAATCCCCTTTCCTCATATGGATTGTTTGTTTCTTCAGACAATTCGTTAGGTCGGTTCCAAAGGTTGATTACTTGACCTTAAGACATGGTTTCGTCATGGCACATCTGGCACCCCACAGCGATCAGAAATTTGACTTTCAAAAATACATAAAACGATCTCTTGAAGAAGATTTCAAGGTGGTGGTCAGCATCAGCCCTCCGATATGGTTCTTTGCTGTCCTCTTCCTACTTTTCAACACCAACGGGTGGAGGGCTTATCTATGGCTACCCTTTGTTCCATTAATTATAGTGTTATTGGTGGGGACAAAGTTGCAAGTGATAATAACGAAGATGGCGCTGAGGATACAAGAAAGAGGAGAAGTGGTGAAAGGAGTACCGGTGGTAGAGCCGGGGGATGACCTCTTTTGGTTCAATCGCCCTCGTCTTATTCTTTACCTTATCAATTTTGTCCTCTTCCAGAATGCTTTTCAGCTTGCCTTTTTTGCTTGGACTTGGAAAGAGTTTGGGATGAAATCTTGTTTCCATGAACACACCGAGGATTTGGTCATCAGAATAACGATGGGAGTCCTCGTTCAAATCCTTTGCAGTTACGTCACATTGCCACTCTATGCTCTAGTCACACAGGTATAG。
2. the application of Rapid identification melon powdery mildew gene described in claim 1, comprising:
1) initiative or the new variety initiative of the breeding material of mildew-resistance MLO gene is carried: utilize the MLO GENE SOURCES given by right 1 to be parent material, in hybridization, backcross progeny, the breeding material that transformation has MLO resistant gene can be obtained.
2) mildew-resistance fundamental research: molecular marker analysis is carried out to the MLO type mildew-resistance gene of right 1; Molecular Mapping or the assignment of genes gene mapping are carried out to this gene; This gene is cloned and Interaction among genes analysis.
3) mildew-resistance transgenic research: utilize the MLO type powdery mildew gene given by claim 1 to carry out transgenic research.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310517585.XA CN104561024A (en) | 2013-10-28 | 2013-10-28 | Rapid identification of MLO (Mycoplasma Like Organism) powdery mildew resistant genes of melon |
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| CN108486272A (en) * | 2018-04-04 | 2018-09-04 | 甘肃省农业科学院蔬菜研究所 | Mildew-resistance germplasm screening method and its application based on muskmelon CmROR2 genes |
| CN112359132A (en) * | 2020-12-07 | 2021-02-12 | 山东省农业科学院蔬菜花卉研究所 | Powdery mildew resistance related SSR molecular marker for muskmelon and application thereof |
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