40 results about "Haynaldia villosa" patented technology

The invention relates to a haynaldia villosa's 6VS chromosome specific molecular marker 6VS-BH1 and an application thereof, and belongs to the technical fields of molecular biology and genetic breeding science. The labeled primer 6VS-BH1 is developed on the basis of wheat-brachypodium distachyon comparative genomics means, and the specific sequence is 6VS-BH1F, as shown in SEQ ID NO.1, and 6VS-BH1R, as shown in SEQ ID NO.2. The molecular marker 6VS-BH1 has the advantages of wide annealing temperature range (60-66 DEG C), good stability, strong product brightness, high resolution and the like. The molecular marker 6VS-BH1, as a co-dominant marker, not only can be used for effectively tracing haynaldia villosa's 6VS chromosome in wheat background, but also can be used for distinguishing a homozygote and heterozygote and for distinguishing a Pm21 gene carried translocation line T6VS.6AL and a PmV gene carried translocation line T6VS.6DL. Therefore, the molecular marker 6VS-BH1 disclosed by the invention has an important practical value in the molecular marker-assisted selection breeding of wheat powdery mildew and the pyramiding breeding of Pm21 gene and PmV gene.

The invention belongs to the field of gene engineering and discloses an NLR gene NLR1-V, an expression vector thereof and application of the NLR gene NLR1-V and the expression vector thereof. A cDNA sequence of the gene NLR1-V with an NLR structural domain is SEQ ID NO.1, and an amino acid sequence encoded by the gene is SEQ ID NO.2. The gene comes from a 6VS chromosome arm of a wheat-haynaldia villosa translocation line 6VS / 6AL Nannong 9918. Inducible expression of NLR1-V by powdery mildew in the powdery mildew resisting wheat variety Nannong 9918 is enhanced. The gene is converted into a susceptible wheat variety Yangmai 158 through transient expression, and the result shows that overexpression of NLR1-V can reduce the haustorium index of the Yangmai 158. Thus, NLR1-V is expected to be used for gene engineering breeding and is expected to improve the powdery mildew resistance of the wheat when introduced to the powdery mildew susceptible wheat variety.

The present invention discloses a KASP molecular marker of a wheat powdery mildew resistance gene Pm21 derived from haynaldia villosa and an application of the KASP molecular marker. The KASP molecular marker is KASP-Pm21 and primers of the KASP molecular marker KASP-Pm21 comprise a haynaldia villosa allelotype upstream primer KASP-Pm21-F, a wheat allelotype upstream primer KASP-Pm21-H and a shared downstream primer KASP-Pm21-C; a nucleotide sequence of the haynaldia villosa allelotype upstream primer KASP-Pm21-F is shown in SEQ ID NO:1; a nucleotide sequence of the wheat allelotype upstream primer KASP-Pm21-H is shown in SEQ ID NO:2; and a nucleoside sequence of the shared downstream primer KASP-Pm21-C is shown in SEQ ID NO:3. The provided KASP molecular marker KASP-Pm21 of the wheat powdery mildew resistance gene Pm21 derived from the haynaldia villosa can realize efficient tracking and detection of the Pm21 gene and is applied to molecular marker assisted selection breeding of the wheat powdery mildew resistance Pm21 gene.

The present invention discloses 3 molecular markers (6SS-1 to 6SS-3) for specifically tracking 6SS chromosomes of aegilops speotoides and a use method of the molecular markers, and relates to the technical field of molecular biology and genetic breeding science. Sequences of primers of the 6SS-1 marker are shown as SEQ ID NO.1 and SEQ ID NO.2, sequences of primers of the 6SS-2 marker are shown asSEQ ID NO.3 and SEQ ID NO.4, and sequences of primers of the 6SS-3 marker are shown as SEQ ID NO.5 and SEQ ID NO.6. The molecular markers can track the Pm12 gene from the aegilops speotoides, also cantrack a Pm21 gene from haynaldia villosa, can simultaneously distinguish and track the Pm12 gene and the Pm21 gene in a pyramiding breeding material, and have important practical value in assistant selection breeding and pyramiding breeding of the two genes.

The invention discloses molecular markers linked with a disease resistance gene Pm21 of wheat and application of the molecular markers in breeding. 25 molecular markers are derived from Triticum aestivum-Haynaldia villosa translocation line NAU427 of the powdery mildew resisting gene Pm21, are tightly linked with the gene Pm21 and can be used for assistant selection of the molecular markers; and selection efficiency and accuracy of disease resisting plants are improved.

The invention discloses a haynaldia villosa calmodulin interacting protein kinase gene and expression vector and application thereof, belonging to the field of gene engineering. The cDNA sequence of HvCIPK1 is SEQ ID NO.1, and the encoded amino acid sequence thereof is SEQ ID No.2. The gene is reported in the haynaldia villosa for the first time, expression is up-regulated due to the induction of powdery mildew germ, and overexpression of the gene in winnow wheat 158 of wheat variety resisting powdery mildew can improve the resistance of the winnow wheat 158 to powdery mildew germ. HvCIPK1 is an important element in disease resistant signal transduction pathway of haynaldia villosa and can be guided into wheat variety resisting powdery mildew, so that the resistance to powdery mildew is improved; the molecular mechanism exploring the action thereof can help to understand the broad-spectrum resistance mechanism of the powdery mildew.

The invention discloses a method for transient expression of a wheat leaf single cell based on efficient gene gun conversion and application of the method. The method for transient expression of the wheat leaf single cell based on the efficient gene gun conversion comprises the following steps of: performing gene gun conversion by using a green fluorescent protein gene as a report gene and a detached leaf of a first wheat leaf as a conversion receptor; immobilizing the wheat leaf by a solid fresh-keeping culture medium, wherein tungsten powder with the diameter of 1 micron is used for gene gun conversion, the using amount is 300ug / gun and the using amount of plasmid is 750ng / gun; and an attacking distance is measured based on 1350psi of cracking membrane helium pressure, a distance between the cracking film and a receptor film is 6 mm, and a distance between the receptor and a stopping net is 6 cm. By the method, haynaldia villosa SGT1 genes are expressed excessively in the detached leaf epidermis cells of the Yangmai 158 of the disease-sensing wheat categories; and a result shows that the index of powdery mildew haustorium in interaction cells can be reduced by the gene transient expression and the invasion of powdery mildew and the formation of the haustorium can be inhibited by the gene so as to increase the resistance of the powdery mildew.

The invention discloses specific molecular markers of haynaldia villosa 4VS chromosome capable of resisting wheat yellow mosaic virus and belongs to the field of biotechnology. The invention firstly discloses two molecular markers CINAU66 and CINAU295 which are capable of tracking the haynaldia villosa 4VS chromosome. In materials containing the haynaldia villosa 4VS chromosome, marker primers CINAU66F / CINAU66R are capable of amplifying about 950bp (base pair) of product; and marker primers CINAU295F / CINAU295R are capable of amplifying about 380bp of product; and the primers are capable of tracking the haynaldia villosa 4VS chromosome. The haynaldia villosa 4VS chromosome capable of resisting the wheat yellow mosaic virus, disclosed by the invention, is capable of proving new gene resources for the breeding of yellow mosaic virus resisting wheat; and the specific molecular markers capable of specifically tracking the haynaldia villosa 4VS chromosome can be used for molecular marker-assisted selective breeding.

The invention belongs to the field of genetic engineering and discloses a haynaldia villosa disulfide isomerase gene and an application thereof. The cDNA (complementary deoxyribonucleic acid) sequence of Hv-PDI (Hantaan Virus-Protein Disulfide Isomerase) is SEQ ID NO. 1, and the encoded amino acid sequence is SEQ ID NO. 2. The gene derives from diploid haynaldia villosa (haynaldia villosa VV, 2n=14), is induced by powdery mildew in powdery mildew-resistant diploid haynaldia villosa, and is greatly expressed. Through transient expression, the gene is transformed to an infected wheat variety Yangmai 158, a result shows that excessive expression of Hv-PDI is capable of reducing the haustorium index of Yangmai 158. Therefore, the Hv-PDI is expectedly used for genetic engineering breeding, and the resistance of wheat to powdery mildew is hopefully improved when the Hv-PDI is introduced to wheat varieties susceptible to powdery mildew.

The invention relates to a haynaldia villosa's 6VS chromosome specific molecular marker 6VS-BH1 and an application thereof, and belongs to the technical fields of molecular biology and genetic breeding science. The labeled primer 6VS-BH1 is developed on the basis of wheat-brachypodium distachyon comparative genomics means, and the specific sequence is 6VS-BH1F, as shown in SEQ ID NO.1, and 6VS-BH1R, as shown in SEQ ID NO.2. The molecular marker 6VS-BH1 has the advantages of wide annealing temperature range (60-66 DEG C), good stability, strong product brightness, high resolution and the like. The molecular marker 6VS-BH1, as a co-dominant marker, not only can be used for effectively tracing haynaldia villosa's 6VS chromosome in wheat background, but also can be used for distinguishing a homozygote and heterozygote and for distinguishing a Pm21 gene carried translocation line T6VS.6AL and a PmV gene carried translocation line T6VS.6DL. Therefore, the molecular marker 6VS-BH1 disclosed by the invention has an important practical value in the molecular marker-assisted selection breeding of wheat powdery mildew and the pyramiding breeding of Pm21 gene and PmV gene.

The invention discloses a complete set of molecular markers for detecting the chromosome arms of Haynaldia villosa and application thereof. The invention provides the complete set of molecular markers for detecting the chromosome arms of Haynaldia villosa, and the complete set of molecular markers are composed of all the or a part of primer pairs (14 primer pairs as shown in sequence 1 to 28 in a sequence table) used for detecting 2V, 3V, 4V, 6V and 7V chromosome arms of Haynaldia villosa. According to the invention, RNA-seq technology is employed for acquiring the transcriptome sequences of Haynaldia villosa; primers are designed on the basis of the transcriptome sequences; and specific markers capable of accurately identifying each chromosome of Haynaldia villosa are screened out, and corresponding amplification primers are designed. Experimental results show that the complete set of primers can realize specific detection of each chromosome of Haynaldia villosa and are successively applied to detection of Haynaldia villosa chromosomes in Triticum durum-Haynaldia villosa amphidiploid and wheat hybrid and backcross generation materials. The complete set of molecular markers in the invention provide a more convenient and effective tool for further research and utilization of Haynaldia villosa.

The invention discloses a kinase gene LeRLK1-V, and an expression vector and application thereof, belonging to the field of genetic engineering. The cDNA sequence of LeRLK1-V is SEQ ID No. 1, and an amino acid sequence coded by LeRLK1-V is SEQ ID No. 2. The gene LeRLK1-V is derived from the 6VS chromosome arm of the Triticum aestivum-Haynaldia villosa translocation line 6VS / 6AL Nannong 9918. LeRLK1-V is enhanced in expression under the induction of powdery mildew in the powdery-mildew-resistant wheat variety Nannong 9918. The gene is used for transforming the susceptible wheat variety Yangmai158 by transient expression, and results show that the overexpression of LeRLK1-V can reduce the haustorium index of Yangmai 158. Therefore, LeRLK1-V is expected to be applied to genetic engineering breeding; and as LeRLK1-V is introduced into wheat varieties susceptible to powdery mildew, the powdery mildew resistance of the wheat varieties is expected to be improved.

The invention provides a molecular marker primer for haynaldia villosa 3V chromosome, namely, a TaGS5-C-1 primer pair and an application of the primer in detecting the haynaldia villosa 3V chromosome. Besides, the invention also provides a PCR method for detecting the haynaldia villosa 3V chromosome on the basis of the primer.