WO2001022804A1 - Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima' - Google Patents

Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima' Download PDF

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Publication number
WO2001022804A1
WO2001022804A1 PCT/IN1999/000051 IN9900051W WO0122804A1 WO 2001022804 A1 WO2001022804 A1 WO 2001022804A1 IN 9900051 W IN9900051 W IN 9900051W WO 0122804 A1 WO0122804 A1 WO 0122804A1
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WO
WIPO (PCT)
Prior art keywords
campestris
polima
cms
crossing
plants
Prior art date
Application number
PCT/IN1999/000051
Other languages
English (en)
Inventor
Jagdish Kumar Verma
Yashpal Singh Sodhi
Neelkantan Arumugam
Arundhati Vukhopadhyay
Akshay Kumar Pradhan
Vibha Gupta
Deepak Pental
Original Assignee
University Of Delhi, South Campus
National Dairy Development Board
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Delhi, South Campus, National Dairy Development Board filed Critical University Of Delhi, South Campus
Priority to PCT/IN1999/000051 priority Critical patent/WO2001022804A1/fr
Priority to CA002385416A priority patent/CA2385416A1/fr
Priority to EP99962497A priority patent/EP1220600A1/fr
Priority to AU18893/00A priority patent/AU1889300A/en
Publication of WO2001022804A1 publication Critical patent/WO2001022804A1/fr
Priority to SE0200962A priority patent/SE0200962D0/xx

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10—Seeds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A01H1/021—Methods of breeding using interspecific crosses, i.e. interspecies crosses
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A01H1/022—Genic fertility modification, e.g. apomixis
    • A01H1/023—Male sterility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/20—Brassicaceae, e.g. canola, broccoli or rucola
    • A01H6/204—Brassica rapa

Definitions

  • the present invention relates to a fertility restorer (Rf) gene for 'Polima' cytoplasmic male sterility (CMS) in Brassica campestris and to a method of producing restorer lines for hybrid seed production.
  • Rf fertility restorer
  • CMS cytoplasmic male sterility
  • breeding for yield improvement in the Brassica crops can be done by utilizing the phenomenon of hybrid vigour or heterosis.
  • two disparate parents having desirable agronomic characters, such as high yield potential, disease resistance, etc. are combined by traditional breeding methods, the Fl hybrid plants show higher yield than either of the parents.
  • One of the most economical and convenient means for producing uniform heterotic population Fl hybrids is to utilize cytoplasmic male sterility (CMS).
  • CMS cytoplasmic male sterility
  • a CMS plant is rendered incapable of producing pollen grains (the male reproductive unit in plants) and therefore cannot get self-fertilized, thus ensuring cross pollination and the development of true hybrid seeds.
  • Factors reasonable for imparting CMS reside in the cytoplasmic organelle the mitrochondria and are transmitted maternally.
  • the Fl hybrid plant obtained by crossing a CMS female parent with a pollen donor parent is sterile and incapable of setting seeds. This is a highly undesirable feature in a crop where seed is the harvested product.
  • this problem is rectified by the presence of a genetic factor in the nucleus of the pollen donor parent. This genetic factor when present in its active form, suppresses the expression of CMS and restores male fertility.
  • This fertility restoration factor (Rf) is an essential component for the production of viable hybrid seeds at commercial scale where seed is the harvested product.
  • Brassica plants a number of CMS systems are available which are reported to have originated either from Brassica species itself or from related genera. For example, 'Polima', one of the well known CMS systems in Brassica has been shown to have its origin in Brassica napus .
  • 'ogura' Another well worked out CMS system in Brassica is 'ogura' which derived its CMS cytoplasm from radish.
  • the restorer factors for the respective CMS systems come from the species that contribute the CMS inducing cytoplasm.
  • the restorer for 'ogura' was transferred from radish to Brassica plants (Delourme et al. 1991, Proc 8th Int. Rapeseed Congr.5, 1506-1510).
  • the restorer for 'Polima' was identified in B. napus itself.
  • Brassica campestris ( synonymus B. rapa) is one of the edible oil seed producing Brassica type plant and is grown in Western Canada, parts of Sweden and Finland and north-west China where summer growing season is too short to accomodate other longer duration Brassica crops.
  • India three distinct types of Brassica campestris, namely brown sarson, yellow sarson and toria are grown in northern and eastern India.
  • significant heterosis for yield in crosses between B. campestris cultivars has been reported from India (Rai and Singh 1994, Indian J genet. 54: 310-314; Varshney and Rao 1997, Indian J. Genet.57: 91-97) and Canada (Schuler et al. 1992, Can J.
  • An object of this invention is to propose a restorer (Rf) gene for the 'Polima' cytoplasmic male sterility in Brassica campestris plants.
  • Another object of this invention is to propose a process for introducing the identified restorer (Rf) gene for the 'Polima' cytoplasmic male sterility in Brassica campestris .
  • a process of producing restorer (Rf) gene for 'Polima' CMS in Brassica campestris (syn. B.rapa) plant which comprises in the steps of : i) Crossing 'Polima' CMS B. napus with B. campestris to produce Fl generation. ii) Subjecting the Fl generation to successive steps of backcrassing with B. campestris and atleast upto
  • a restorer factor for 'Polima' CMS in Brassica campestris there is provided the identification of a restorer factor for 'Polima' CMS in Brassica campestris.
  • the process also provides that the fertility restoration is governed by a single Mendelian locus.
  • 'Polima' CMS was transferred from B. napus var. ISN 706 to B. campestris. To effect this transfer, interspecific cross was made between 'Polima' CMS B. napus var. ISN 706 with B. campestris plants. The resultant Fl hybrid plant was repeatedly back-crossed to normal
  • the BC3 generation B. campestris containing 'Polima' CMS cytoplasm was subsequently crossed with other B. campeatris varieties such as long duration canola quality 'Candle' or accession 'ATC 94211'.
  • the Fl plants segregated for male fertility and male sterility.
  • the fertile Fl plants were self-pollinated.
  • the resultant F2 progeny plants were scored for the segregation of male fertile and male sterile plants in order to establish the inheritance pattern of the fertility restorer factor.
  • Fig.l is a schematic flow diagram showing the breeding methodology for the identification of restorer lines for 'Polima' CMS B. campestris. It comprises the prepa- ration of Fl hybrid by making inter-specific cross between 'Polima' CMS B. napus var. ISN 706 and B. campestris var. 'Pusa kalyani ' followed by three back- crosses of the said Fl hybrid to B. campestris var. 'Pusa kalyani'. Subsequently, BC3 'Polima' CMS B. cam- pestris var. 'Pusa kalyani' was crossed with 'Candle' and 'ATC 94211'. Fertile Fl plants were self pollinated to raise F2 progeny.
  • the present invention relates to a plant of the species B. campestris, characterised in that it contains a
  • Example of Brassica campestris plant usable in the present method include any plant line or cultivar that includes the Rf factor for the 'Polima'
  • the 'Polima' CMS B. campestris is initially developed through conventional breeding method by crossing 'Polima' CMS B. napus var. ISN 706 with a B. campestris plant.
  • a breeding process for producing a stable 'Polima' CMS line in B. campestris which comprises in the steps of :
  • B. napus var, ISN 706 Making an inter-specific cross between 'Polima' B. napus var, ISN 706 with B. campestris to produce Fl generation.
  • Normal fertile B. napus ISN 706 is a synthe- tic Brassica napus the seeds of which were obtained from Indian Agricultural Research Institute, New Delhi, India.
  • 'Polima' CMS was introduced in B. napus ISN 706 from a unknown French variety containing 'Polima' cytoplasm ( Sodhi et al. 1993, Plant Breeding 110: 334- 337).
  • siliqua seeds
  • the siliqua were collected, examined for the seeds and the seeds were planted.
  • the resulting Fl plants were male sterile and partially female fertile and hence, upon back-crossing to B. campestris produced a few seeds. The seeds were planted to raise BCl generation.
  • B. campestris variety used in the present invention is, for example, B. campestris var. 'Pusa kalyani'.
  • B. campestris var. 'Pusa kalyani' is a released cultivar in India.
  • B. campestris var. 'Pusa kalyani' BC3 is crossed with two other lines of B. campestris namely, 'Candle' and 'ATC 94211'.
  • the Fl plants of 'Polima' 0 'Pusa kalyani' x 'Candle' and 'Polima' 'Pusa kalyani' x 'ATC 94211' segregated for male fertility and male sterility.
  • the fertile Fl plants had normal floral features with respect to flower opening and floral organ development.
  • the pollen viability of the male 5 fertile Fl plants of these two crosses are 99% and the production of the viable pollen are similar to that of normal B. campestris var.
  • 'Candle' is a Canadian 'Canola* quality cultivar having low erucic acid and low glucosino- late.
  • the seeds of 'candle' were obtained from Agriculture Canada, Saskatoon, Canada.
  • 'ATC94211' is an Australian low erucic acid line and seeds were obtained from Brassica germplasm bank from Australia.
  • the present invention involves conventional field breeding techniques.
  • the breeding experiments were conducted twice in a year, utilizing the winter growing season (October- April) in the plains (Delhi) and summer growing season (May-September) in the higher altitude areas of Northern Himalayas (Leh, J&K) .
  • the plants were grown by sowing seeds in 3m long rows with 40cm spacing between rows and 15cm spacing between plants within the rows.
  • Fl seeds were planted in the next growing season. Germination occurred within 5-7 days. Standard intercultural practices were followed. Plants were intermediate between B. napus and B. campestris in their morphology and were uniform in appearance. The plants flowered after 70 days and flowering was uniform. All the plants were male sterile and partially female sterile. Flowers had short stamens bearing conical whitish anthers, which were devoid of pollen grains.
  • the said Fl plants were back-crossed with B. campestris var. 'Pusa kalyani'. About 100 crosses were made, each cross comprising about 10-12 buds. Siliqua developed in situ. The development was however retarded as compared to that of normal siliqua and only 2-3 siliqua developed per cross (1.5-2.5cm having 2-3 seeds as against around 5.0cm and 16-18 seeds in normal case).
  • BCl seeds were harvested and sown in the next growing season. Plants showed variation in their morphological features. Some were intermediate between B. napus and B. campestris and some resembled B. campestris more closely.
  • BC2 seeds were harvested and sown in the next growing season. Resemblance of the plants to B. campestris was more perceptible in this generation and majority of them came to flowe- ring within 60 days of sowing. Flowers were completely male sterile and female fertility was normal. About 15 crosses were made each having 10-12 buds. Siliqua development was almost normal, with each siliqua having on an average 12 seeds.
  • the BC3 seeds were harvested and sown during next growing season. The plants completely resembled B. campestris var. 'Pusa kalyani' phenotypically and came to flowering within 60 days. The flowers were completely male sterile with crinkled petals, short stamens and small whitish conical anthers.
  • the Fl seeds were space planted in eight rows at a distance of approximately 25cm between plants for each of the crosses.
  • Fl plants derived from both 'Pusa kalyani' x 'Candle' and 'Pusa kalyani' x 'ATC 94211' displayed unit phenotype and floral morphology intermediate between the respective parents, The plants showed 1:1 segregation for male fertility and sterility.
  • the pollen viability of the male fertile Fl plants of these two crosses are 99% and the production of the viable pollen are similar to that of normal B. campestris var. 'Pusa kalyani' (Table 1).
  • the Fl plants showed normal seed set under open pollination.
  • the fertile Fl plants were selfed.
  • the inflorescence Prior to the opening of the buds the inflorescence were covered securely with butter paper bags to prevent cross-pollination.
  • the siliqua were allowed to develop within the confine of the bag till the grain filling stage. On an average, 12 seeds per siliqua were obtained. The selfed seeds were harvested.
  • F2 seeds of each cross were space planted with a distance of about 20cm between plants in 22 rows each. Segregation was observed for plant morphology and flowering duration. At the time of flowering plants were scored in the field for male fertility and male sterility. The segregation pattern was recorded as 3 male fertile: 1 male sterile. Plants were female sterile since seed setting was normal on open pollination.

Abstract

La présente invention concerne un gène restaurateur pour la stérilité mâle cytoplasmique 'Polima' dans des plantes Brassica camprestris (synonymus Brassica rapa), et un procédé d'obtention de lignes restauratrices dans Brassica campestris. Cette invention a révélé par ailleurs que la restauration de la stérilité mâle cytoplasmique 'Polima' dans B. campestris est commandée par un gène dominant unique. L'identification d'un gène restaurateur de la stérilité mâle cytoplasmique 'Polima' présente une utilité potentielle pour la mise au point de variétés hybrides de B. campestris.
PCT/IN1999/000051 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima' WO2001022804A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
PCT/IN1999/000051 WO2001022804A1 (fr) 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima'
CA002385416A CA2385416A1 (fr) 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique "polima"
EP99962497A EP1220600A1 (fr) 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique "polima"
AU18893/00A AU1889300A (en) 1999-09-28 1999-09-28 Fertility restorer gene for "polima" cytoplasmic male sterility
SE0200962A SE0200962D0 (sv) 1999-09-28 2002-03-28 Fertilitetsbevarande gen för "polima"-cytoplasmatisk hansterilitet

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/IN1999/000051 WO2001022804A1 (fr) 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima'

Publications (1)

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WO2001022804A1 true WO2001022804A1 (fr) 2001-04-05

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PCT/IN1999/000051 WO2001022804A1 (fr) 1999-09-28 1999-09-28 Gene de restauration de la fertilite dans le cas de sterilite male cytoplasmique 'polima'

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EP (1) EP1220600A1 (fr)
AU (1) AU1889300A (fr)
CA (1) CA2385416A1 (fr)
SE (1) SE0200962D0 (fr)
WO (1) WO2001022804A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006021972A1 (fr) * 2004-08-26 2006-03-02 Dhara Vegetable Oil And Foods Company Limited Nouveau système de stérilité cytoplasmique pour espèces de brassicées et utilisation pour production de graines hybrides de moutarde indienne brassica juncea à base de graines oléagineuses
CN111154908A (zh) * 2020-01-20 2020-05-15 中国农业科学院油料作物研究所 用于鉴别芸薹属作物Cam和Pol胞质类型的KASP标记引物及其应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110786233B (zh) * 2019-12-11 2022-09-23 贵州省油菜研究所 用核三系油菜f2代分离选育同质临保及纯合不育系方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0267753A1 (fr) * 1986-11-07 1988-05-18 Allelix Inc. Méthode pour le transfert d'éléments cytoplasmiques en Brassica
EP0599042A1 (fr) * 1992-10-14 1994-06-01 Mitsubishi Corporation Procédé pour l'introduction d'un gène restaurant la fertilité et pour le production de plantes F1-hybrides de Brassica
WO1997002737A1 (fr) * 1995-07-07 1997-01-30 Zenco (No.4) Limited Hybrides de colza 'canola' ayant un systeme de sterilite male cytoplasmique

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0267753A1 (fr) * 1986-11-07 1988-05-18 Allelix Inc. Méthode pour le transfert d'éléments cytoplasmiques en Brassica
EP0599042A1 (fr) * 1992-10-14 1994-06-01 Mitsubishi Corporation Procédé pour l'introduction d'un gène restaurant la fertilité et pour le production de plantes F1-hybrides de Brassica
WO1997002737A1 (fr) * 1995-07-07 1997-01-30 Zenco (No.4) Limited Hybrides de colza 'canola' ayant un systeme de sterilite male cytoplasmique

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006021972A1 (fr) * 2004-08-26 2006-03-02 Dhara Vegetable Oil And Foods Company Limited Nouveau système de stérilité cytoplasmique pour espèces de brassicées et utilisation pour production de graines hybrides de moutarde indienne brassica juncea à base de graines oléagineuses
US8030548B2 (en) 2004-08-26 2011-10-04 Dhara Vegetable Oil And Foods Company Limited Cytoplasmic male sterility system for Brassica species and its use for hybrid seed production in indian oilseed mustard Brassica juncea
CN111154908A (zh) * 2020-01-20 2020-05-15 中国农业科学院油料作物研究所 用于鉴别芸薹属作物Cam和Pol胞质类型的KASP标记引物及其应用

Also Published As

Publication number Publication date
CA2385416A1 (fr) 2001-04-05
EP1220600A1 (fr) 2002-07-10
AU1889300A (en) 2001-04-30
SE0200962D0 (sv) 2002-03-28

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