CN102703463A - Rapid identification of powdery mildew resistance Lotus corniculatus genes utilizing comparative genomics - Google Patents
Rapid identification of powdery mildew resistance Lotus corniculatus genes utilizing comparative genomics Download PDFInfo
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Abstract
The invention relates to rapid identification of powdery mildew resistance Lotus corniculatus genes utilizing comparative genomics, relates to knowledge of subjects such as plant comparative genomics, genetics and bioinformatics, and belongs to the technical field of plant biotechnology. The rapid identification of powdery mildew resistance Lotus corniculatus genes utilizing the comparative genomics mainly includes the steps of firstly, downloading full genomic sequences of Lotus corniculatus and collecting MLO (mildew resistance locus o) genes; secondly, identifying the MLO genes; thirdly, identifying phylogenetic relationship of the MLO genes; and fourthly comparing the MLO powdery mildew resistance genes. By the rapid identification, mining cycle of the powdery mildew resistance genes is shortened, and the powdery mildew resistance genes can be identified quickly. Corresponding co-separation functional marks (SNP (single nucleotide polymorphism), Scar and the like) can be developed through candidate powdery mildew resistance genes identified, the rapid identification is also available for molecular marker-assisted selection of the powdery mildew resistance genes, and accuracy is high. The rapid identification can also be used with other molecular markers for resistance genes to create multiresistance breeding materials, and accordingly breeding period is shortened, breeding efficiency is improved, and basis for elaborating powdery mildew resistance Lotus corniculatus molecular mechanisms is laid.
Description
Technical field
The present invention is by means of Root or stem of Littleleaf Indianmulberry order-checking whole genome sequence; Utilize method Rapid identification Root or stem of Littleleaf Indianmulberry powdery mildew genes such as plant comparative genomics, genetics, information biology and candidate gene strategy, be mainly concerned with the download of Root or stem of Littleleaf Indianmulberry whole genome sequence, the evaluation of candidate gene; The comparison of gene; Means such as cluster, and then identify powdery mildew gene, belong to the Plant Biotechnology scientific domain.
Background technology
Root or stem of Littleleaf Indianmulberry (
Lotus corniculatus) have another name called five leaves grass (Herba Galii Bungei), bird foot beans, Birdfoot; It is perennial pulse family Lotus herbage; Originate in area, temperate zone, Eurasia; On China Hebei, Yunnan, Guizhou, Sichuan, Gansu and other places distribution is arranged all, Root or stem of Littleleaf Indianmulberry is a kind of forage grass that is widely used in orchard ground cover, green manure, grassland improvement, also is a kind of Chinese medicinal plant that can be clearing heat and detoxicating.Powdery Mildew is a disease common on the Root or stem of Littleleaf Indianmulberry, and the Root or stem of Littleleaf Indianmulberry growing area has generation more in China, influences forage yield and quality.The pathogenic bacteria that causes this type disease be the pea powdery mildew (
Erysiphe pisiDC.), different name knotweed white powder germ, general living powdery mildew clover specialized form belong to a kind of fungi of Ascomycotina.Except parasitism on Root or stem of Littleleaf Indianmulberry, this type of germ is also parasitic on leguminous plantss such as Trifolium, Melilotus suaveolens Ledeb. genus, Medicago, flat Mu Macroptilium and pea.Pathogenic bacteria in the season of growth pea powdery mildew infect with conidium, conidium borrows wind-force to propagate distance quite far away.Conidium quantity is big, under optimum conditions, causes disease popular very soon.Sunshine, abundance was windy, and soil and atmospheric moisture are medium, and the more high environment of height above sea level helps this sick generation.
White powder germ infringement Root or stem of Littleleaf Indianmulberry can make position such as the two sides, stem, petiole, pod of plant leaf be injured, and wherein infecting object is main with blade.White mustiness spot appears in disease portion surface, and the initial stage scab is little and rounded, and the back enlarges and converges mutually, so that covers big of blade to the full wafer leaflet.Had a strong impact on the normal growth and development process of Root or stem of Littleleaf Indianmulberry plant, the great financial loss that can already bring for herbage.The utilization of disease-resistant genetic resources and breeding for disease resistance are the effective meanss of the control of plant disease of generally acknowledging, the clone of mildew-resistance gene and obtain to help further developing of Root or stem of Littleleaf Indianmulberry breeding research.
MLO type disease-resistant gene is one type of special disease-resistant gene of plant.The investigator find the earliest MLO (
MIldew resistance
lOcus
o ) the gene pairs powder mildew resistance is to start from 1937-1938, gathered the barley of a lot of kinds by the fritz in Ethiopia, two strains systems wherein to the white powder germ (
Blumeria graminisF. sp.
Hordei) physiological strains that all oneself know all have efficient resistance.Further research shows, the recessive mutation mlo of MLO gene can make barley that the physiological strain of nearly all oneself knowledge barley powdery mildew bacteria is produced resistance lasting, wide spectrum in the barley.Recently, the investigator finds that the mildew-resistance gene of a lot of plants all is a MLO type Gene Handling, like tomato, and pea, Arabidopis thaliana, capsicum, rose, Root or stem of Littleleaf Indianmulberry or the like.Therefore breeding has important effect to the plant mildew-resistance to excavate MLO type disease-resistant gene in the plant.
At present, the disease-resistant gene method commonly used of excavating commonly used has map based cloning, methods such as transposon tagging.But because the fundamental research of Root or stem of Littleleaf Indianmulberry is not deep enough, not only the time is long but also very difficult these genes of cloning exactly therefore to utilize these methods.Therefore, how the MLO type disease-resistant gene in the Rapid identification Root or stem of Littleleaf Indianmulberry will become the important prerequisite of Root or stem of Littleleaf Indianmulberry mildew-resistance breeding.
Plant comparative genomics (Comparative Genomics) is based on Genome Atlas and the order-checking basis, and known gene and genome structure are compared, and understands the subject of function, expression mechanism and the spore of gene.Utilize between model plant genome and other Plant Genome on the coded sequence and structural homology, clone's other plant gene discloses gene function and molecular mechanism, illustrates spore relation and genomic immanent structure.Method that this patent adopted and thinking: the research of model plant arabidopsis gene group has disclosed the function of MLO type gene; Utilize the homology clone Root or stem of Littleleaf Indianmulberry MLO type disease-resistant gene on its order of gene; According to the characteristics of meliority on the model plant Arabidopis thaliana experimental system and known MLO type disease-resistant gene, quick " seizure " Root or stem of Littleleaf Indianmulberry mildew-resistance gene.In recent years, the completion of Root or stem of Littleleaf Indianmulberry gene order-checking provides condition for our fast mining Root or stem of Littleleaf Indianmulberry powdery mildew gene.It is prerequisite that this patent has been introduced with the Root or stem of Littleleaf Indianmulberry whole genome sequence, in conjunction with knowledge such as comparative genomics, genetics, genomics, information biology and candidate gene strategy, fast mining powdery mildew gene.
Summary of the invention
Technical problem
The purpose of this invention is to provide a kind of through combining knowledge such as plant comparative genomics, plant genetics, genomics and information biology, fast mining Root or stem of Littleleaf Indianmulberry mildew-resistance gene.Its result can be used for the exploitation of Root or stem of Littleleaf Indianmulberry powdery mildew gene compact linkage molecule mark on the one hand, carries out the molecular marker assisted selection breeding, also for other crop powdery mildew gene identification reference frame is provided on the other hand.
Technical scheme
Cardinal principle: first plant mildew-resistance gene (MLO) is cloned from barley, discovers that this gene is one type of special disease-resistant gene, is different from most of NBS (nucleotide-binding site) type disease-resistant gene of previous clone; Subsequently, the investigator has cloned powdery mildew gene in succession from plants such as tomato, Arabidopis thaliana, pea, capsicum, Root or stem of Littleleaf Indianmulberry, and what discover these genes encodings all is MLO type disease-resistant gene.Subsequently, numerous investigators confirm that through test of many times MLO type disease-resistant gene has become one type of mildew-resistance gene of plant specific.Find that further plant MLO type gene is a gene family; And the MLO gene family that derives from different plant species is carried out the Phylogenetic Relationships analysis finds, in the different plant species mildew-resistance gene always cluster become one type together, this type MLO gene all has the characteristic feature of mildew-resistance gene sequence.The completion of Root or stem of Littleleaf Indianmulberry gene order-checking provides a convenient for excavating powdery mildew gene.Therefore, can identify the Root or stem of Littleleaf Indianmulberry powdery mildew gene by means of the Phylogenetic Relationships of MLO gene family in the full genome of the Root or stem of Littleleaf Indianmulberry of having checked order and the MLO powdery mildew gene of having cloned and for the amino acid conservative property of keeping powdery mildew gene MLO critical function.
Key step is following:
1) collection of the download of Root or stem of Littleleaf Indianmulberry whole genome sequence and MLO type gene thereof
At first download the Root or stem of Littleleaf Indianmulberry whole genome sequence from Root or stem of Littleleaf Indianmulberry sequenced genes group DB (http://www.kazusa.or.jp/lotus/index.html); Use " DNATOOLS " software that the full genome amino acid sequence data of Root or stem of Littleleaf Indianmulberry that obtains is set up DB; Use pfam DB (protein family DB then; Http:// pfam.janelia.org/search/sequence) hidden Markov model in (HMM) carries out Blastp (E-value=0.001) sequence alignment to the aminoacid sequence of MLO structural domain with the full genome amino acid sequence database of having set up of Root or stem of Littleleaf Indianmulberry, and preliminary screening goes out candidate gene sequence.Secondly, utilize the MLO type gene order of having announced, the Root or stem of Littleleaf Indianmulberry genome database is carried out the BLAST comparison, obtain candidate gene sequence.
2) evaluation of Root or stem of Littleleaf Indianmulberry MLO type gene family
With the candidate gene of the homologous nucleotide sequence that obtains in the The above results, analyze through Pfam (E-value=1.0), remove the gene order (Fig. 1) of not having ' MLO ' structural domain.The ClustalW instrument that again candidate's disease-resistant gene sequence is provided through MEGA3.1 software (multisequencing comparison program) carries out the multisequencing comparison, removes Tumor-necrosis factor glycoproteins.
3) Phylogenetic Relationships through plant MLO type gene is identified candidate's Root or stem of Littleleaf Indianmulberry MLO type powdery mildew gene
Because previous research is verified; Dicotyledons MLO type powdery mildew gene is positioned at the same district of plant MLO Phylogenetic Tree group; Therefore in Phylogenetic Relationships research; We are MLO type mildew-resistance gene and the cluster analysis together of Root or stem of Littleleaf Indianmulberry MLO type gene of the MLO type gene family of Arabidopis thaliana and some other crops, to obtain candidate's Root or stem of Littleleaf Indianmulberry mildew-resistance gene (Fig. 2).
4) comparison of Root or stem of Littleleaf Indianmulberry powdery mildew gene and known plant MLO powdery mildew gene
Utilize BioXM 2.6 softwares the aminoacid sequence of the MLO powdery mildew gene of Root or stem of Littleleaf Indianmulberry candidate's MLO type powdery mildew gene and Arabidopis thaliana, tomato, pea, barley to be converted to the file of Fasta form; These files are imported BioEdit 7.0 softwares; Use in this software Clustal software to carry out the multisequencing comparison, disclose the conservative property in powdery mildew gene important amino acid residue and zone.Thereby further identify Root or stem of Littleleaf Indianmulberry candidate's powdery mildew gene (Fig. 3).
Positively effect of the present invention:
1) shortens the Root or stem of Littleleaf Indianmulberry powdery mildew gene and excavated the cycle, helped the Rapid identification of powdery mildew gene.Employing ordinary method (map based cloning, transposon tagging etc.) excavation mildew-resistance gene not only takes time and effort, efficient is low, and is difficult to successfully.The present invention is based on plant comparative genomics, genetics, bioinformatics method fast mining Root or stem of Littleleaf Indianmulberry powdery mildew gene, not only can shorten the time, can also improve powdery mildew gene and identify efficient.
2) Root or stem of Littleleaf Indianmulberry (
Lotus cornioulatus L.) belong to the pulse family Lotus, be research leguminous crop pattern species.Because the Root or stem of Littleleaf Indianmulberry hereditary basis is narrow, the germ plasm resource variety is low, therefore identifies relatively difficulty of Root or stem of Littleleaf Indianmulberry powdery mildew gene through conventional molecule marker (RAPD, ISSR, SSR, AFLP etc.).The candidate's powdery mildew gene exploitation that is tested and appraised is divided into from functional mark (SNP, SCAR etc.) accordingly, can be used for the molecular marker assisted selection of disease-resistant gene fast, and accuracy is high.
3) initiative of multiresistance breeding material.Based on the functional molecular marker of the powdery mildew gene of new evaluation exploitation, combine the molecule marker of localized other disease-resistant genes, carry out the initiative of multiresistance breeding material, can shortening the breeding cycle, the raising breeding efficiency.
4) lay a good foundation for setting forth Root or stem of Littleleaf Indianmulberry mildew-resistance molecular mechanism.The evaluation of Root or stem of Littleleaf Indianmulberry mildew-resistance gene; Gene silencing (virus induced gene silencing through transgenic technology, RNAi, virus induction; VIGS) molecular mechanism of research mildew-resistance such as technology provides genetic resources, helps setting forth fast the mechanism of action of Root or stem of Littleleaf Indianmulberry mildew-resistance.
Description of drawings
The evaluation of Fig. 1 Root or stem of Littleleaf Indianmulberry MLO gene;
That this figure shows is 10 MLO type gene identification results, and each gene all contains ' MLO ' conserved domain.
The evaluation of the Phylogenetic Relationships analysis of Fig. 2 plant MLO gene family and Root or stem of Littleleaf Indianmulberry MLO type powdery mildew gene thereof;
Arabidopis thaliana is the model plant of plant science research; Grow in the tree at constructing system; The powdery mildew gene (PsMLO) of 15 MLO type genes of Arabidopis thaliana (wherein 3 genes are powdery mildew genes: AtMLO02, AtMLO06 and AtMLO12), tomato mildew-resistance gene (SlMLO), barley powdery mildew gene (HvMLO and HvMLO02) and pea is selected and is used for and the cluster analysis of Root or stem of Littleleaf Indianmulberry MLO type gene.Identify 1 Root or stem of Littleleaf Indianmulberry candidate's MLO type powdery mildew gene altogether.The gene of italic mark is exactly a candidate Root or stem of Littleleaf Indianmulberry powdery mildew gene among the figure.
The compare of analysis of Fig. 3 Root or stem of Littleleaf Indianmulberry MLO type powdery mildew gene;
1 Root or stem of Littleleaf Indianmulberry powdery mildew gene and barley (HvMLO), tomato (SlMLO), pea (PsMLO), Arabidopis thaliana powdery mildew gene (AtMLO02; AtMLO06 and AtMLO12) compare, identify that powdery mildew infects the amino-acid residue and regional conservative type that plays an important role.TM1-TM7 representes 7 revolving die zones of Root or stem of Littleleaf Indianmulberry MLO type powdery mildew gene among the figure; The black round dot representes that powdery mildew infects important amino-acid residue; CaMBD representes the calmodulin land; I and II represent powdery mildew is infected important amino acid region.
Embodiment
The evaluation of disease-resistant gene has important effect in research of crop disease-resistant theory of heredity and disease-resistant variety seed selection.Present method can Rapid identification go out the Root or stem of Littleleaf Indianmulberry powdery mildew gene.The practical implementation process is following:
1) collection and the evaluation of Root or stem of Littleleaf Indianmulberry MLO type gene
In order to obtain the whole MLO type gene family member of Root or stem of Littleleaf Indianmulberry; We are at first with the MLO type gene of Arabidopis thaliana; The mildew-resistance MLO gene order of tomato, pea, capsicum, rose, Root or stem of Littleleaf Indianmulberry makes up the HMM model, from the Root or stem of Littleleaf Indianmulberry genome sequence, receives rope MLO type gene; Secondly with the MLO gene order delivered in the Different Crop as target sequence (from DFCI DB: 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 DB: AY967408, AF384145, AF384144; AY029312-AY029315, AY029317-AY029319, Z95352; AF369563-AF369565, AF369567, AF369569-AF369576; Z83834, Z95496, AY581255); Root or stem of Littleleaf Indianmulberry DB (http://www.kazusa.or.jp/lotus/index.html) is carried out the BLAST comparison, select the highest sequence of similarity to download, obtained 10 candidates' MLO type gene (LjT43M12.190.r2.a altogether; Chr1.CM0017.1150.r2.a; Chr6.CM0066.530.r2.a; Chr6.CM0114.350.r2.m; Chr1.CM.0113.1360.r2.d; LiT40B16.20.r2.m; Chr6.CM0508.460.r2.m; Chr2.CM0272.1030.r2.d; Chr1.CM0113.1440.r2.d; LjSGA_047574.1).
2) evaluation of Root or stem of Littleleaf Indianmulberry MLO type gene family
In order further to verify these MLO gene accuracys, we have carried out the evaluation of conserved domain " MLO " to these 10 candidates' MLO gene.Aminoacid sequence with each candidate's MLO type gene is a benchmark, on PFAM (http://pfam.sanger.ac.uk/) website, carries out the evaluation of ' MLO ' conserved domain, and concrete outcome is seen Fig. 1.
3) the Phylogenetic Relationships analysis of Root or stem of Littleleaf Indianmulberry MLO type gene
In the research formerly, find that the dicotyledons powdery mildew gene aggregates into district's group; Therefore; Grow in the tree at constructing system; We have selected the powdery mildew gene and the Root or stem of Littleleaf Indianmulberry MLO type gene cluster analysis cluster analysis together of 15 MLO type genes (wherein 3 genes are powdery mildew genes: AtMLO02, AtMLO06 and AtMLO12), tomato mildew-resistance gene, barley powdery mildew gene and the pea of model plant Arabidopis thaliana.Root or stem of Littleleaf Indianmulberry MLO type gene and other crop powdery mildew gene protein sequences are carried out the multisequencing couplet join (adopting Clustal X 1.83 softwares to carry out), and utilize Genedoc software (http://www.nrbsc.org/gfx/genedoc/index.html) to show that multisequencing joins the result who joins.The Clustal multisequencing is joined the result who joins output in MEGA 4.0 softwares, and (neighbor-joining NJ), utilizes the Bootstrapping method that these evolutionary trees are assessed to utilize this software to make up the adjacency tree respectively.The result finds in dicotyledons mildew-resistance gene district group, to have 1 Root or stem of Littleleaf Indianmulberry candidate's MLO type powdery mildew gene (see figure 2).
4) comparison of Root or stem of Littleleaf Indianmulberry MLO type disease-resistant gene
In the research of barley MLO type powdery mildew gene, found some important areas and single amino acids in the research in succession, they infect big wheat powdery mildew has irreplaceable effect.In the Root or stem of Littleleaf Indianmulberry powdery mildew gene of identifying 1 candidate; Whether these important areas and amino acid high conservative, and we have carried out compare of analysis to 3 mildew-resistance genes (AtMLO02, AtMLO6 and AtMLO12), tomato powdery mildew gene (SlMLO), the powdery mildew of pea gene (PsMLO) from Arabidopis thaliana.1 candidate's of discovery Root or stem of Littleleaf Indianmulberry powdery mildew gene and 7 of known MLO type powdery mildew genes are striden the film district, 30 important amino acid, 1 calmodulin land (CaMBD) and two important zone (I and II) high conservatives (Fig. 3).
Claims (4)
1. Root or stem of Littleleaf Indianmulberry mildew-resistance gene is characterized in that being selected from following 1 gene:
LjT40B16.20.r2.m
Aminoacid sequence:
MAKEDRVYERTLEETPTWAVAVVCFVLLAISIIIEHIIHAIGKWFKKKRKNALFEALEMVKGELMLMGFISLLLTVLQDPISKICISKSVASTWHPCANPKTKTKHDADSDSDDSNHRKLLEYFDLIPRRVLATKGYDKCHDKGKVALVSAYGIHQLHIFIFVLAIFHILQCIITLGLGRIKMRRWTRWEDETKTFEYQFYNDPERFRFARDTTFGRRHLSTWARSPVSLWIVSFLRQFYGSVTKVDYMALRHGFVMAHLAPGNDANFNFQKYINRSLEEDFKVVVGISPIIWFCAVLFLLTNAHGWYSYYWLPFIPLIIILLVGTKLQMIITMMGLRIQDRGEVIKGAPMVEPGDHLFWFNRPSLLLFLIHLVLFQNAFQLAFFSWSTYEFSLRSCFHEKNVDIIIRLTLGVVIQFLCSYVTLPLYALVTQMGSTLKPTIFNERVADALKKWHHTAKRQVKHSKHSEANSITPFSSRPATPTHGMSPVHLLHRHLAGRSDSVETSPRTSNYENEQWDVEGSNSPSNHPTGVDEIQLEVLEQRSEATELPITSTRHEISIAVSDFSFQKRHTGSD
Nucleotide sequence:
ATGGCTAAGGAAGACAGAGTTTATGAAAGAACTCTGGAAGAAACACCAACCTGGGCTGTTGCAGTTGTGTGCTTTGTGCTGCTTGCTATTTCAATCATAATTGAACATATTATTCATGCTATTGGAAAGTGGTTCAAAAAGAAGCGCAAAAATGCTCTTTTTGAAGCATTGGAAATGGTCAAAGGAGAGCTTATGCTGATGGGATTCATATCCTTGCTACTAACTGTGCTCCAAGATCCAATTTCTAAGATCTGTATATCAAAAAGTGTTGCATCCACTTGGCATCCTTGTGCCAATCCAAAGACAAAGACTAAACATGATGCAGATTCTGACAGTGATGATTCCAATCATAGGAAACTTCTCGAATATTTTGACCTCATTCCGCGGCGTGTTCTAGCTACAAAAGGATATGACAAGTGCCATGATAAGGGAAAAGTTGCCCTTGTTTCTGCATATGGGATCCACCAGCTCCACATATTTATTTTTGTGTTAGCTATTTTTCATATCCTACAATGCATTATAACACTTGGTTTGGGAAGGATTAAGATGAGACGCTGGACGAGATGGGAAGATGAAACAAAGACTTTTGAATATCAATTCTATAATGATCCTGAGAGGTTCAGATTTGCAAGGGACACAACATTTGGACGAAGGCACTTGAGCACGTGGGCTCGCTCACCAGTTTCCTTATGGATAGTTAGCTTCCTCAGACAATTCTATGGATCTGTTACTAAAGTTGATTACATGGCATTACGGCATGGATTTGTCATGGCACATCTAGCACCAGGAAACGATGCAAATTTCAATTTCCAGAAGTATATCAATAGATCACTAGAGGAGGATTTTAAAGTTGTGGTGGGAATAAGCCCTATTATATGGTTCTGTGCGGTGCTATTTCTGCTGACTAATGCTCATGGGTGGTATTCTTATTATTGGCTTCCATTTATCCCCTTAATTATAATCTTGCTAGTGGGTACTAAGCTACAAATGATCATAACAATGATGGGTTTGAGGATTCAAGACAGAGGTGAAGTAATCAAGGGTGCACCTATGGTTGAGCCAGGTGATCATCTGTTCTGGTTCAATCGTCCCAGCCTCCTTCTCTTTTTGATTCATCTTGTTCTCTTTCAGAATGCCTTTCAACTAGCTTTCTTTTCTTGGAGTACATATGAGTTCTCTCTAAGGTCATGCTTCCACGAAAAAAATGTGGATATTATCATTAGACTTACATTGGGGGTTGTAATACAATTCCTATGCAGCTATGTCACTTTGCCTCTTTATGCTCTAGTTACACAGATGGGTTCAACCTTGAAACCAACAATTTTCAATGAAAGAGTAGCAGATGCACTGAAGAAATGGCACCACACAGCCAAAAGGCAAGTCAAACACAGCAAGCATTCTGAGGCTAACAGCATCACACCATTCTCGAGCAGGCCAGCGACGCCAACGCACGGCATGTCTCCGGTTCACCTGCTGCACAGACACCTTGCCGGCCGCAGCGACAGTGTGGAAACTTCTCCGAGGACATCGAACTACGAGAATGAGCAGTGGGATGTTGAAGGCTCAAATTCTCCAAGTAACCACCCAACAGGAGTTGATGAGATACAGTTGGAAGTTTTGGAGCAACGTTCAGAAGCAACAGAATTGCCTATTACCAGCACTAGACATGAAATCAGCATTGCTGTATCCGATTTTTCTTTCCAGAAGCGACACACCGGTAGTGATTAG。
2. the application of the said Rapid identification Root or stem of Littleleaf Indianmulberry of claim 1 powdery mildew gene comprises:
1) initiative of breeding material.
3.2) breeding practice of mildew-resistance.
4.3) mildew-resistance fundamental research.
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| CN104561034A (en) * | 2013-10-28 | 2015-04-29 | 常熟市杜桥稻米专业合作社 | Rapid identification method for powdery mildew resistant gene of foxtail millet |
| CN104561032A (en) * | 2013-10-28 | 2015-04-29 | 常熟市董浜镇里睦蔬菜专业合作社 | Quick identification of powdery mildew resistant gene for barley |
| CN104561255A (en) * | 2013-10-28 | 2015-04-29 | 常熟市杜桥稻米专业合作社 | Rapid identification on MLO type anti-disease gene of rice |
| CN104561253A (en) * | 2013-10-28 | 2015-04-29 | 南农大(常熟)新农村发展研究院有限公司 | Rapid identification of corn powdery mildew resistant genes |
| CN104561033A (en) * | 2013-10-28 | 2015-04-29 | 常熟市杜桥稻米专业合作社 | Rapid identification on wheat MLO type powdery mildew gene |
| CN104561254A (en) * | 2013-10-28 | 2015-04-29 | 常熟市董浜镇里睦蔬菜专业合作社 | Rapid identification method of powdery mildew-resistant genes of cabbage |
| CN104561024A (en) * | 2013-10-28 | 2015-04-29 | 南农大(常熟)新农村发展研究院有限公司 | Rapid identification of MLO (Mycoplasma Like Organism) powdery mildew resistant genes of melon |
| CN104593480A (en) * | 2013-10-30 | 2015-05-06 | 江苏省常熟现代农业产业园区发展有限公司 | Application of comparative genomics to rapid identification of phaseolus vulgaris mildew resistance locus o gene |
| CN104593379A (en) * | 2013-10-30 | 2015-05-06 | 江苏省常熟现代农业产业园区发展有限公司 | Rapid identification of powdery mildew resistant gene of Chinese cabbage by using comparative genomics |
| CN104593481A (en) * | 2013-10-30 | 2015-05-06 | 江苏省常熟现代农业产业园区发展有限公司 | Rapid identification of soybean anti-powdery mildew gene by using candidate gene strategy |
| CN104611407A (en) * | 2013-11-04 | 2015-05-13 | 常熟市董浜镇北港蔬菜专业合作社 | Rapid identification of strawberry powdery mildew gene by using comparative genomics |
| CN104611409A (en) * | 2013-11-04 | 2015-05-13 | 常熟市董浜镇北港蔬菜专业合作社 | Rapid identification of Capsella bursa-pastoris anti-powdery mildew gene by using comparative genomics |
| CN104611408A (en) * | 2013-11-04 | 2015-05-13 | 常熟市董浜镇北港蔬菜专业合作社 | Rapid identification of watermelon MLO type anti-powdery mildew gene |
| CN113832244A (en) * | 2021-09-03 | 2021-12-24 | 武汉市农业科学院 | MNP (MNP) core primer combination for molecular identification of lotus DNA (deoxyribonucleic acid) varieties and application of MNP core primer combination |
| CN113832244B (en) * | 2021-09-03 | 2024-05-24 | 武汉市农业科学院 | MNP core primer combination for lotus DNA variety molecular identification and application thereof |
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