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PRODUCTION METHOD FOR TRANSFORMATION-SENSITIVE BARLEY

Foreign code F180009491
File No. S2016-0868-C0
Posted date Nov 1, 2018
Country WIPO
International application number 2017JP023218
International publication number WO 2017222051
Date of international filing Jun 23, 2017
Date of international publication Dec 28, 2017
Priority data
  • P2016-124749 (Jun 23, 2016) JP
Title PRODUCTION METHOD FOR TRANSFORMATION-SENSITIVE BARLEY
Abstract By measuring allele segregation distortion in an F2 group obtained by crossing Golden Promise and Haruna Nijo, the inventors of the present invention were the first in the world to identify the genomic region that is the basis for sensitivity to Agrobacterium-mediated transformation in barley. Using the existence of said genomic region as an index, the inventors discovered that it is possible to efficiently produce barley that is sensitive to Agrobacterium-mediated transformation.
Outline of related art and contending technology BACKGROUND ART
Barley (Hordeum vulgare) is grown, a well-characterized 5.1Gbp diploid genome of a cereal. Barley is, the amount of grain in the world of production in a first rank and at 4, food, beer brewing, and an animal feed are used. From the importance of the crop, many agriculturally or barley genetic characteristics of the industry has been analyzed. Transformation, and an essential tool in genetics, DNA into the genome of the organism by introducing, as well as the introduction of a new transformation, the analysis of endogenous gene becomes possible. Unfortunately, barley in the genetic analysis, transformation and restricted by a technical problem for, such as Arabidopsis thaliana or other plant species is rice transformed by Agrobacterium method in stark contrast to the ease of the. Barley Agrob.alpha method is widely used in the transformation method, and developed by Tingay et al. (Non-Patent Document 1), the golden as explants utilizes immature embryos of the promise. Other than this as a transformation technique of the barley, pollen iguri few alternative culture (Igri) is one of the (non-patent document 2). Research purposes of the characteristic analysis and expressing the golden promise iguri alleles can limit, in this situation, a great challenge in barley leads to the complementary analysis. In general, the analysis of functional alleles, via a plurality of golden promise backcrossed, while leaving the target allele, the genome region of the other line that is replaced with the golden promise type needs to be developed. Recently, Yeo et al. (Non-Patent Document 3) is, the hook portion and a non-host with respect to the plant analysis system suitable for the function of the resistance of the barley to develop, the golden promise suspret transformation inherited from the CPU and the ability of the disease (SusPtrit) having both high sensitivity (Golden SusPtrit) golden suspret barley for the new developed system. Recombinant in the system, the enhancement of the efficiency of the transformed Agrobacterium method, and the transformation efficiency of the promise of golden view and indicative of the presence of genetic factors. In this study, in the transformed Agrobacterium method of the golden promise becomes a factor in order to predict the genetic region, a population of doubled haploid the same mapping (SG062N) transforming the non-sensitive strain sensitive system (SG133N) comparison of the differences between the genetic (non-patent document 3) is performed.
Transformation specific genetic factors of the capability itself is not investigated, barley green callus in view of the chute 4 and for reproduction of one of the quantitative trait locus (QTLs) is, the previous, and (Morex) stephen molex (Steptoe) identified in the hybrid (non-patent document 4, 5). Cogan et al. (Brassica oleracea) brassica oleracea (non-patent document 6) is in, using seeds ofplants, Agrobacterium, transformation of the root using rhizogenes 3 involved one QTL was identified. 3 One of the two loci 2, these areas overlap each other generated by the genome in the vicinity of the paralogs is assumed that the (non-patent document 7). And potato (Solanum tuberosum) in, (non-patent document 8) El-Kharbotly et al. is, R1 genes (resistance to bacteria Vital Phytophthora infestans and potato) is linked to the, transformation efficiency was found that the loci involved. In the case of rice, Nishimura et al. (Non-Patent Document 9) is, the major QTL (PSR) in the promoter of the chute reproduction involved in the nitrite reductase (ferredoxin-nitrate reductase) ferredoxin (NiR) isolated the gene encoding. In this case, the callus from the seeds ofplants, the bevel region or of the complete genomic gene NiR of bolus over-expressed by introducing a cDNA, rice KOSHIHIKARI of the reproducing efficiency is increased. Interestingly, Tyagi et al. (Non-Patent Document 10) is, the reproduction of the green and the overshoot of the QTL on chromosome 6H according to the barley, (AK371794) present in the vicinity of the locus reported.
The efficiency of the transformed Agrobacterium method, the type of plant material are investigated not only in the configuration or a genetic, environmental and technical factors depend greatly. Technical factors, culture conditions, media composition, Agrobacterium strain type, and the particular binary and comprises a selection marker is employed (Non-Patent Document 11). Associated with the interaction of the genetic factors Agrob.alpha (T-DNA host for incorporation into the genome, the selection conditions of the cell division, and from the callus and the reproduction of the view of the genes) as well as, the type of the explants used for transformation also an important role (non-patent document 11).
Recently, (Trans Activator-Like Effector Nucleases) or TALENs CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated proteins 9) such as a plant genome editing techniques have been developed. However, the utility of these techniques in barley, Lawrenson Budhagatapalli et al. (Non-Patent Document 12) (Non-Patent Document 13) and reported by the inventors as, in most plant species, the first step of introducing these nuclease transformed as necessary by the fact that the constraint has been received.
Scope of claims (In Japanese)請求の範囲
[請求項1]
 ゴールデンプロミスと任意のオオムギ品種を交雑させ、得られた後代において、下記(a)~(c)の少なくとも1つのゲノム領域の存在を検出し、当該ゲノム領域がホモまたはヘテロでゴールデンプロミス型である個体を選抜することを含む、アグロバクテリウム法による形質転換に対して感受性のオオムギの作出方法。
 (a)染色体3HにおけるマーカーNIASHv1109O03_00000798_3HとマーカーNIASHv1150F05_00000946_3Hで挟まれたゲノム領域
 (b)染色体2HにおけるマーカーNIASHv2027F11_00000300_2HとマーカーNIASHv2016D11_00000337_2Hで挟まれたゲノム領域
 (c)染色体2HにおけるマーカーNIASHv2108A23_00000279_2HとマーカーFLOUbaf102a14_00001505_2Hで挟まれたゲノム領域
[請求項2]
 ゴールデンプロミスと任意のオオムギ品種を交雑させ、得られた後代において、下記(a)~(c)の少なくとも1つのマーカーの存在を検出し、当該マーカーがホモまたはヘテロでゴールデンプロミス型である個体を選抜することを含む、アグロバクテリウム法による形質転換に対して感受性のオオムギの作出方法。
 (a)染色体3HにおけるマーカーNIASHv1109O03_00000798_3H、マーカーNIASHv1150F05_00000946_3H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
 (b)染色体2HにおけるマーカーNIASHv2027F11_00000300_2H、マーカーNIASHv2016D11_00000337_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
 (c)染色体2HにおけるマーカーNIASHv2108A23_00000279_2H、マーカーFLOUbaf102a14_00001505_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
[請求項3]
 下記(a)~(c)の少なくとも1つのマーカーを挟み込むように設計された一対のオリゴヌクレオチドを含む、オオムギにおいてアグロバクテリウム法による形質転換に対する感受性を検査するための試薬。
 (a)染色体3HにおけるマーカーNIASHv1109O03_00000798_3H、マーカーNIASHv1150F05_00000946_3H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカーからなる群より選択されるマーカー
 (b)染色体2HにおけるマーカーNIASHv2027F11_00000300_2H、マーカーNIASHv2016D11_00000337_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカー
 (c)染色体2HにおけるマーカーNIASHv2108A23_00000279_2H、マーカーFLOUbaf102a14_00001505_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
[請求項4]
 下記(a)~(c)の少なくとも1つのマーカーを含むゲノム上の塩基配列にハイブリダイズするオリゴヌクレオチドを含む、オオムギにおいてアグロバクテリウム法による形質転換に対する感受性を検査するための試薬。
 (a)染色体3HにおけるマーカーNIASHv1109O03_00000798_3H、マーカーNIASHv1150F05_00000946_3H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
 (b)染色体2HにおけるマーカーNIASHv2027F11_00000300_2H、マーカーNIASHv2016D11_00000337_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
 (c)染色体2HにおけるマーカーNIASHv2108A23_00000279_2H、マーカーFLOUbaf102a14_00001505_2H、およびそれらマーカーで挟まれたゲノム領域に存在するマーカーからなる群より選択されるマーカー
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • NATIONAL UNIVERSITY CORPORATION OKAYAMA UNIVERSITY
  • Inventor
  • SATO Kazuhiro
  • HISANO Hiroshi
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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