Top > Search of International Patents > RICE RT-TYPE CYTOPLASMIC MALE STERILITY CAUSAL GENE AND USE THEREOF

RICE RT-TYPE CYTOPLASMIC MALE STERILITY CAUSAL GENE AND USE THEREOF

Foreign code F150008360
Posted date Jun 29, 2015
Country WIPO
International application number 2013JP066528
International publication number WO 2014027502
Date of international filing Jun 10, 2013
Date of international publication Feb 20, 2014
Priority data
  • P2012-180824 (Aug 17, 2012) JP
Title RICE RT-TYPE CYTOPLASMIC MALE STERILITY CAUSAL GENE AND USE THEREOF
Abstract The purpose of the present invention is to identify the causal gene of rice RT-type cytoplasmic male sterility and to provide a means for determining a rice sterile line that possesses RT-type male-sterile cytoplasm by using said causal gene as a DNA marker, in order to employ rice RT-type cytoplasmic male sterility in a three-line method. The present invention provides: a causal gene of rice RT-type cytoplasmic male sterility that includes a DNA comprising a base sequence represented by SEQ ID NO: 1 or 3; and a method for determining a rice sterile line that possesses RT-type male-sterile cytoplasm by detecting the presence of said gene in a rice to be tested.
Outline of related art and contending technology BACKGROUND ART
One generation hybrid breeding method is also referred to as hybrid cultivar breeding method, in accordance with the parents have excellent traits, and, in the hybrid varieties can be grown for bias, used in a variety of plant. Hybrid rice in rice (one margin hybrid varieties) for breeding, cytoplasmic male sterility (CMS: Cytoplasmic Male Sterility) of three-line system is used. Cytoplasmic male sterility is, by the operation of the abnormal mitochondrial gene, a phenomenon that interfere with the formation of pollen. To the mitochondrion of the cytoplasmic male sterility gene responsible for male sterility is present, on the other hand, the fertility restoration gene with a nuclear-encoded modified chimeric gene product is performed to avoid the inhibition believed. Is a three-line system, carrying cytoplasmic male sterility system for the 'male sterile line', carrying a fertility restoration gene with a single system (Rf) 'fertility recovery system', and, nuclear genes is identical to the male sterile line which do not have cytoplasmic male sterile system for the 'maintain system' refers to using breeding methods. Using these 3 system, the recovery system (i) pollen from male sterile line by mating of the hybrid seed can be obtained, (ii) maintaining the male sterile line by crossing with pollen of the system to maintain the male sterile line, are able to grow. In the three-line system utilizing cytoplasmic male sterility can be carried out, cytoplasmic male sterility reliably whether the rice male sterile line to hold the assay is required. Conventional, plant cytoplasmic male sterility in order to determine the genotype of, first, and positive and negative test crossing and self-pollinated, seed formation rate to investigate the frequency of appearance of individuals 0% is necessary. However, this method requires much time and year required much labor is required, in addition, the temperature and duration of sunshine seed fertility may easily affected by environmental conditions such as the assay cannot be accurately. Therefore, efficiently rapid and easy identification of cytoplasmic male sterility rice is a method for assaying has been desired. Compared with a conventional three-line system hybrid rice varieties using the rice variety is grown Chinsurah Boro II India BT-type cytoplasmic male sterility or wild rice varieties of hainan island WA-type cytoplasmic male sterility has been used, the main flow WA-type cytoplasmic male sterility (non-patent document 1) in. From the viewpoint of the total rice cultivation area of the world, the interrupt 1 single WA-type male sterility cytoplasm becomes dependent on the. Up to now WA-type cytoplasmic male sterility rice cytoplasmic male sterility is the cause of orf126 as a candidate for the communication of the other genes have been reported (non-patent document 2). However, cytoplasmic male sterility is directly involved in or has not been demonstrated experimentally. Therefore, WA-type cytoplasmic male sterility, cytoplasmic causal genes, fertility restoration gene with fertility of the identity of the recovery system is clearly not, molecular basis remains unsolved is being used. In addition, other than the above cytoplasmic male sterility for the system to provide a report some rice. For example, type RT98 (RT98A) cytoplasmic male sterility system is of the university of the ryukyus motomura et al., wild rice Oryza rufipogon K98 system (a system number National Laboratory W1109) and 1 times the parent, #8 in table 65 and the three consecutive backcross can be performed in a system which has grown, but apparently normal pollen, has no function of fertilization. In addition, the growth in the course, the same cytoplasmic but that RT98A, K98 having a fertility restoration gene derived from fertile fertility is restored in order to grow a RT98C recovery system (non-patent document 3) are. In addition, type RT102 (RT102A) cytoplasmic male sterility system is, of the system K102 in the same manner as Oryza rufipogon (National Laboratory system number W1125) to a parent 1 times, 8 times in one continuous backcross Nos 65 performed in a system which has grown, and the pollen does not accumulate in the starch accumulation and was observed in a mixed with, both not being capable of fertilization. In addition, the growth of course, is the same with the cytoplasm RT102A, having a fertility restoration gene derived from K102 for regenerating the recovery system RT102C is restored fertility are growing (non-patent document 4). However, type RT98 and type RT102 cytoplasmic male sterility cytoplasmic male sterility is the cause of the mitochondrial gene has not been reported. In order to use the growth of hybrid varieties, these RT-type cytoplasmic male sterility to identify genes that cause mitochondrial, cloning has been desired.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 以下の(a)~(c)のいずれかに示すDNAを含むイネRT型細胞質雄性不稔性の原因遺伝子。
 (a)配列番号1または3に示す塩基配列からなるDNA
 (b)配列番号1または3に示す塩基配列からなるDNAと相補的な塩基配列からなるDNAとストリンジェントな条件下でハイブリダイズし、かつイネRT型細胞質雄性不稔性に関与するDNA
 (c)配列番号1または3に示す塩基配列に対して80%以上の相同性を有する塩基配列からなり、かつイネRT型細胞質雄性不稔性に関与するDNA

[請求項2]
 以下の(d)~(f)のいずれかに示すタンパク質をコードするイネRT型細胞質雄性不稔性の原因遺伝子。
 (d)配列番号2または4に示すアミノ酸配列からなるタンパク質
 (e)配列番号2または4に示すアミノ酸配列において1若しくは数個のアミノ酸が欠失、置換若しくは付加されたアミノ酸配列からなり、かつイネRT型細胞質雄性不稔性に関与するタンパク質
 (f)配列番号2または4に示すアミノ酸配列に対して80%以上の相同性を有するアミノ酸配列からなり、かつイネRT型細胞質雄性不稔性に関与するタンパク質

[請求項3]
 被検イネにおける請求項1または2に記載の遺伝子の存在を検出することを特徴とする、RT型雄性不稔細胞質を有するイネ系統の判別方法。

[請求項4]
 請求項1または2に記載の遺伝子の存在の検出が、配列番号1もしくは配列番号3に記載の塩基配列、または該塩基配列の部分配列を含むDNA領域を検出することにより行われる、請求項3に記載の方法。

[請求項5]
 検出が、PCR法またはハイブリダイゼーション法により行われる、請求項3または4に記載の方法。

[請求項6]
 請求項3~5のいずれかに記載の方法により判別されたRT型雄性不稔細胞質を有するイネ系統を受粉系統とし、該細胞質雄性不稔に対して花粉稔性を回復する稔性回復遺伝子を保有または導入したイネ系統を授粉系統として交配することを特徴とするイネハイブリッド種子の生産方法。

[請求項7]
 配列番号1もしくは配列番号3に記載の塩基配列、または該塩基配列の部分配列を増幅するための、少なくとも15塩基以上の長さのオリゴヌクレオチドから構成される、RT型細胞質雄性不稔性の原因遺伝子増幅用プライマーセット。

[請求項8]
 以下の(A)または(B)のプライマーセットである、請求項7に記載のプライマーセット。
(A)配列番号5に示す塩基配列を含むオリゴヌクレオチドと配列番号6に示す塩基配列を含むオリゴヌクレオチドとから構成される、RT98型細胞質雄性不稔性の原因遺伝子増幅用プライマーセット
(B)配列番号7に示す塩基配列を含むオリゴヌクレオチドと配列番号8に示す塩基配列を含むオリゴヌクレオチドとから構成される、RT102型細胞質雄性不稔性の原因遺伝子増幅用プライマーセット

[請求項9]
 配列番号1もしくは配列番号3に記載の塩基配列、または該塩基配列の部分配列にハイブリダイズし、請求項1に記載の遺伝子を検出するための少なくとも15塩基以上の長さのオリゴヌクレオチドからなるプローブ。

[請求項10]
 請求項7または8に記載のプライマーセット、および/または、請求項9に記載のプローブを含む、RT型雄性不稔細胞質を有するイネ系統の判別用キット。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • TOHOKU UNIVERSITY
  • UNIVERSITY OF THE RYUKYUS
  • Inventor
  • TORIYAMA, KINYA
  • KAZAMA, TOMOHIKO
  • MOTOMURA, KEIJI
  • IGARASHI, KEISUKE
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 DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KN KP KR KZ LA LC LK LR LS LT 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 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 TD TG
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