WO2000016610A1 - Procede permettant d'induire une organogenese directe et in vitro dans les oignons - Google Patents

Procede permettant d'induire une organogenese directe et in vitro dans les oignons Download PDF

Info

Publication number
WO2000016610A1
WO2000016610A1 PCT/SI1999/000022 SI9900022W WO0016610A1 WO 2000016610 A1 WO2000016610 A1 WO 2000016610A1 SI 9900022 W SI9900022 W SI 9900022W WO 0016610 A1 WO0016610 A1 WO 0016610A1
Authority
WO
WIPO (PCT)
Prior art keywords
media
induction
differentiation
contain
process according
Prior art date
Application number
PCT/SI1999/000022
Other languages
English (en)
Inventor
Borut Bohanec
Zlata Luthar
Original Assignee
Univerza V Ljubljani, Biotehniška Fakulteta
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 Univerza V Ljubljani, Biotehniška Fakulteta filed Critical Univerza V Ljubljani, Biotehniška Fakulteta
Priority to AU57685/99A priority Critical patent/AU5768599A/en
Publication of WO2000016610A1 publication Critical patent/WO2000016610A1/fr

Links

Classifications

    • 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
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • 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
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/002—Culture media for tissue culture

Definitions

  • This invention refers to a field of plant biotechnology, specifically to a new process for the induction of direct in vitro organogenesis in onion, with a specific application use in micropropagation or genetic transformation in onion.
  • Onion (Allium cepa L.) is the second most important vegetable species worldwide and is produced in almost all climatic regions. It can be multiplied by seeds, sets or vegetatively. Vegetative in vitro methods are used for multiplication of valuable breeding lines including maintaining male sterile lines, used in hybrid seed production. Methods of direct in vitro organogenesis can be further used for successful genetic transformation in onion.
  • the claimed invention is a process for induction of direct in vitro organogenesis in onion, comprising the steps of:
  • the applied differentiation and induction media contain as solidifiers gellan-gum, mixture of gellan-gum and agar, or only agar.
  • the duration of the growth on induction media is from 3 to 12 days.
  • the induction and differentiation media contain sucrose, glucose or maltose as a source of carbohydrates.
  • the induction and/or differentiation media contain 25-100 g/1 of sucrose.
  • the induction media contain 2,4-dichlorophenoxyacetic acid or picloram as a sources of auxins.
  • the induction media contain auxin or auxin and 6-benzylaminopurine, thidiazuron or isopentenyladenin (2ip) as sources of cytokinins.
  • the differentiation media contain thidiazuron or 6-benzylaminopurine as sources of cytokinins.
  • the induction and differentiation steps are performed in light or in darkness.
  • composition of macro and micro elements in induction and differentiation media corresponds to BDS media prepared according to Dunstan and Short, 1977, B5 media
  • the plant material used in the following Examples originated from different, publicly available sources, cultivars were received from genebanks or were purchased at retail, and inbred lines were received from the US public breeding program (Dr. M.J. Havey, USDA, Madison, Wisconsin, USA).
  • Various genotypes of onion were used in these experiments: Belokranjka (Slovenia), Ptujska rdeca (Slovenia), Stuttgarter Riesen, Timor, Shenshu Yellow, Yamaguchi Koudaka, Texas Early Grano 502, experimental hybrid XPH 3371 FI (Asgrow), hybrids 70723 (B1717BxB2923B) and 70719 (B2371CxB2923B), inbred lines B2355B, B2923B, MSU2935B, MSU5718B, MSU8155B.
  • flower buds were cultured in Petri dishes 100 mm in diameter (30 per dish) on induction media described later. The flowers were on induction media for 6 days (where not said otherwise). After the induction period, flowers were subcultured on Petri dishes containing differentiation media, as described in the following paragraphs.
  • the embodiment 2 differed from embodiment 1 in that the flowers were cultured on induction media and were extracted (perianth removed) before transfer to differentiation media ovaries.
  • Petri dishes were sealed with ParafilmTM (American National Can, Greenwich, CT, USA) to prevent evaporation and exposed to a 16/8 hours photoperiod at 21-23 °C and illumination of approximately 80 ⁇ mol m "2 s " '.
  • ParafilmTM American National Can, Greenwich, CT, USA
  • Such shoots were divided and subcultured on the elongation media, on which they elongated and produced normal plantlets. A proportion of these organogenic structures remained as nodular bumps. When such clusters were subcultured on hormone free media, elongation of shoots occurred, although the organogenic potential was preserved for at least 3 subcultures.
  • the most suitable medium for elongation of shoots was basal medium (or half strength basal medium) excluding phytohormones, with the concentration of sucrose or glucose lowered (20-70g/l, recommendable concentration 30g/l).
  • Basal medium or half strength basal medium
  • concentration of sucrose or glucose lowered (20-70g/l, recommendable concentration 30g/l.
  • the media could additionally contain auxins such as 0.5-1.0 mg/1 indolebutyric acid (IBA).
  • IBA indolebutyric acid
  • the duration of the induction stage had a significant effect on regeneration.
  • the highest shoot regeneration was observed on ovaries that were placed on differentiation media after 6 days. Shorter or longer induction treatment resulted in decreased regeneration. At 6 days, the lowest hyperhydration also occurred.
  • sucrose concentration in induction and differentiation media was studied using 3 genotypes and embodiment 2.
  • Three different concentrations of sucrose used were: 100 g/l (Il/Dl), 50 g/l (I4/D4) and 25 g/l (I5/D5).
  • the results are presented in Table 4.
  • auxin composition in induction media was studied.
  • Induction media contained 2 mg/1 2,4 D (14), 1 mg/1 picloram (18), 2 mg/1 picloram (19) or 5 mg/1 NAA (110).
  • the differentiation media were Dl and D4. The results are presented in Table 5.
  • Induction media contained 2 mg/l BAP (14), cytokinin omitted (11 1), 1 mg/1 TDZ (112) and 2 mg/1 2ip (113).
  • the differentiation medium was D4. The results are presented in Table 6.
  • the highest regeneration was obtained on medium 112, the difference was statistically significant compared to 111 and 113 (first genotype), with the second genotype the highest regeneration was obtained on 14 medium.
  • the percentage of hyperhydrated shoots was also low on 112 medium. Omitting cytokinin in the induction media (111) greatly reduced regeneration.
  • Induction and differentiation media included BDS medium (I4/D4), B5 medium (I14/D12) or MS medium (I15/D13). The results are presented in Table 9. Table 9:

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Cette invention concerne un procédé permettant d'induire une organogenèse directe et in vitro dans les oignons tout en omettant la phase de cals de l'oignon, cette organogenèse étant déclenchée à partir des organes reproducteurs, notamment les fleurs ou les ovaires. Les tissus embryonnaires formés lors du processus de régénération peuvent être utilisés afin de procéder à d'autres transformations génétiques. L'organogenèse somatique directe est un procédé qui offre un niveau élevé de propagation clonale (micro-propagation) à partir de plantes donneuses individuelles.
PCT/SI1999/000022 1998-09-24 1999-09-22 Procede permettant d'induire une organogenese directe et in vitro dans les oignons WO2000016610A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU57685/99A AU5768599A (en) 1998-09-24 1999-09-22 A process for the induction of direct (in vitro) organogenesis in onion

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SIP-9800247 1998-09-24
SI9800247A SI20053A (sl) 1998-09-24 1998-09-24 Postopek za indukcijo neposredne in vitro organogeneze pri čebuli

Publications (1)

Publication Number Publication Date
WO2000016610A1 true WO2000016610A1 (fr) 2000-03-30

Family

ID=20432337

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SI1999/000022 WO2000016610A1 (fr) 1998-09-24 1999-09-22 Procede permettant d'induire une organogenese directe et in vitro dans les oignons

Country Status (3)

Country Link
AU (1) AU5768599A (fr)
SI (1) SI20053A (fr)
WO (1) WO2000016610A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6995016B2 (en) 2000-08-17 2006-02-07 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture And Agri-Food Process for inducing direct somatic embryogenesis in immature scutella cells of pooideae, and rapidly regenerating fertile plants
CN102870674A (zh) * 2012-09-05 2013-01-16 广州白云华南生物科技有限公司 一种红葱组培快速繁殖方法
EP2925739A4 (fr) * 2012-11-28 2016-07-27 Stichting Dienst Landbouwkundi Dihydropyridines substituées pour l'embryogenèse somatique dans des plantes
CN110226517A (zh) * 2019-06-26 2019-09-13 北京市农林科学院 一种洋葱离体再生的方法及其使用的培养基
CN117441617A (zh) * 2023-12-22 2024-01-26 北京花乡花木集团有限公司 一种北葱的组织培养方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0814166A2 (fr) * 1989-08-09 1997-12-29 DeKalb Genetics Corporation Méthode et compositions pour la préparation de plantes monocotylédones fertiles ainsi que leurs cellules transformées de manière stabile

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0814166A2 (fr) * 1989-08-09 1997-12-29 DeKalb Genetics Corporation Méthode et compositions pour la préparation de plantes monocotylédones fertiles ainsi que leurs cellules transformées de manière stabile

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
COHAT, J.: "Obtention chez l'échalote (Allium cepa L var aggregantum) de plantes haploïdes gynogénétiques par culture in vitro de boutons floraux", AGRONOMIE, vol. 14, 1994, pages 299 - 304, XP000879486 *
DATABASE BIOSIS BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1992, CAMPION, B. ET AL.: "Advances in haploid plant induction in onion (Allium cepa L.) through in vitro gynogenesis", XP002131955 *
DATABASE BIOSIS BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1995, BOHANEC, B.: "Studies of gynogenesis in onion (Allium cepa L.): induction procedures and genetic analysis of regenerants.", XP002131954 *
DATABASE BIOSIS BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1996, JAKSE, M. ET AL.: "Effect of media components on the gynogenic regeneration of onion (Allium cepa L.) cultivars and analysis of regenerants.", XP002131953 *
DATABASE CAB CAB INTERNATIONAL, WALLINGFORD, OXON, GB; 1997, JEONG HAEBOONG ET AL.: "Plant redifferentiation and in vitro multiplication of onion by shoot primodium culture", XP002131956 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6995016B2 (en) 2000-08-17 2006-02-07 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture And Agri-Food Process for inducing direct somatic embryogenesis in immature scutella cells of pooideae, and rapidly regenerating fertile plants
CN102870674A (zh) * 2012-09-05 2013-01-16 广州白云华南生物科技有限公司 一种红葱组培快速繁殖方法
CN102870674B (zh) * 2012-09-05 2013-12-25 广州白云华南生物科技有限公司 一种红葱组培快速繁殖方法
EP2925739A4 (fr) * 2012-11-28 2016-07-27 Stichting Dienst Landbouwkundi Dihydropyridines substituées pour l'embryogenèse somatique dans des plantes
CN110226517A (zh) * 2019-06-26 2019-09-13 北京市农林科学院 一种洋葱离体再生的方法及其使用的培养基
CN110226517B (zh) * 2019-06-26 2021-06-01 北京市农林科学院 一种洋葱离体再生的方法及其使用的培养基
CN117441617A (zh) * 2023-12-22 2024-01-26 北京花乡花木集团有限公司 一种北葱的组织培养方法
CN117441617B (zh) * 2023-12-22 2024-04-02 北京花乡花木集团有限公司 一种北葱的组织培养方法

Also Published As

Publication number Publication date
SI20053A (sl) 2000-04-30
AU5768599A (en) 2000-04-10

Similar Documents

Publication Publication Date Title
Chalupa Plant regeneration by somatic embryogenesis from cultured immature embryos of oak (Querem robur L.) and linden (Tilia cordata Mill.)
Hazra et al. Direct somatic embryogenesis in peanut (Arachis hypogea)
US5856191A (en) Method for regeneration of coniferous plants by somatic embryogenesis in culture media containing abscisic acid
Luthar et al. Induction of direct somatic organogenesis in onion (Allium cepa L.) using a two-step flower or ovary culture
Kim et al. Somatic embryogenesis and plant regeneration from immature zygotic embryos of Japanese larch (Larix leptolepis)
Gladfelter et al. De novo shoot organogenesis of Pinus eldarica Medw. in vitro: I. Reproducible regeneration from long-term callus cultures
Tilkat et al. Micropropagation of mature male pistachio Pistacia vera L.
Hossain et al. High efficiency plant regeneration from petiole explants of Carica papaya L. through organogenesis
Zhang et al. Direct organogenesis and plantlet regeneration from mature zygotic embryos of masson pine (Pinus massoniana L.)
Hita et al. Somatic embryogenesis from chickpea (Cicer arietinum L.) inmature cotyledons: The effect of zeatin, gibberellic acid and indole-3-butyric acid
Alttaher et al. High-frequency induction of multiple shoots and plant regeneration from cotyledonary node explants of tongkat ali (Eurycoma longifolia jack).
WO2000016610A1 (fr) Procede permettant d'induire une organogenese directe et in vitro dans les oignons
WO1993012645A1 (fr) Embryogenese somatique et regeneration vegetale du cacao
EP2332405B1 (fr) Procédés et milieux de culture tissulaires destinés à induire une maturation de l'embryon somatique de conifère
CN110896854B (zh) 一种促进油松体细胞胚胎发育成熟的培养方法
JP3198895B2 (ja) ユーカリプタス・グロブラスのクローン増殖方法
Supaibulwatana et al. Organogenesis and somatic embryogenesis from young flower buds of Agapanthus africanus Hoffmanns
US7598073B2 (en) Methods for producing high yields of zygotic-like cotyledonary pine embryos utilizing media that include a disaccharide and glucose
Manohari et al. Efficient plant regeneration in small cardamom (Elettaria cardamomum Maton.) through somatic embryogenesis
CN110896853A (zh) 一种促进枫香属植物体胚成熟的液体悬浮培养方法
Yokoya et al. Regeneration of rose plants from cell and tissue cultures
Vujović et al. Improvement of in vitro micropropagation of black currant'Čačanska Crna'
Farsi et al. Open the windows toward somatic embryogenesis of leaf explants of persian walnut (Juglans regia L.)
Shintiavira et al. In vitro organogenesis of Lisianthus [Eustoma grandiflorum (Raf.) Shinn] derived from leaf explant.
Sudhersan et al. Somatic embryogenesis and production of synthetic seeds in Ziziphus

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase