Influences of aging and cloning methods on the capacity for somatic embryogenesis of a mature Hevea brasiliensis genotype

BackgroundRubber tree (Hevea brasiliensis) laticifers are the source of natural rubber. Rubber production depends on endogenous and exogenous ethylene (ethephon). AP2/ERF transcription factors, and especially Ethylene-Response Factors, play a crucial role in plant development and response to biotic and abiotic stresses. This study set out to sequence transcript expressed in various tissues using next-generation sequencing and to identify AP2/ERF superfamily in the rubber tree.ResultsThe 454 sequencing technique was used to produce five tissue-type transcript libraries (leaf, bark, latex, embryogenic tissues and root). Reads from all libraries were pooled and reassembled to improve mRNA lengths and produce a global library. One hundred and seventy-three AP2/ERF contigs were identified by in silico analysis based on the amino acid sequence of the conserved AP2 domain from the global library. The 142 contigs with the full AP2 domain were classified into three main families (20 AP2 members, 115 ERF members divided into 11 groups, and 4 RAV members) and 3 soloist members. Fifty-nine AP2/ERF transcripts were found in latex. Alongside the microRNA172 already described in plants, eleven additional microRNAs were predicted to inhibit Hevea AP2/ERF transcripts.ConclusionsHevea has a similar number of AP2/ERF genes to that of other dicot species. We adapted the alignment and classification methods to data from next-generation sequencing techniques to provide reliable information. We observed several specific features for the ERF family. Three HbSoloist members form a group in Hevea. Several AP2/ERF genes highly expressed in latex suggest they have a specific function in Hevea. The analysis of AP2/ERF transcripts in Hevea presented here provides the basis for studying the molecular regulation of latex production in response to abiotic stresses and latex cell differentiation.

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“…Self-rooted plants were produced by somatic embryogenesis at the CIRAD laboratory [55,80]. Total mRNAs were isolated from different tissues.…”

Section: Methodsmentioning

confidence: 99%

Identification of the Hevea brasiliensisAP2/ERF superfamily by RNA sequencing

et al. 2013

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“…A second method was developed from friable callus maintained over the long term with a view to large-scale multiplication. This method has evolved with the use of fragments of embryos derived from SEP in order to rapidly establish embryogenic friable callus lines [29,32]. Lastly, cryopreservation of friable calli has been incorporated into the process to limit tissue proliferation and, thereby, the risks linked to somaclonal variation [33].…”

Section: Introductionmentioning

confidence: 99%

Some ethylene biosynthesis and AP2/ERF genes reveal a specific pattern of expression during somatic embryogenesis in Hevea brasiliensis

et al. 2012

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BackgroundEthylene production and signalling play an important role in somatic embryogenesis, especially for species that are recalcitrant in in vitro culture. The AP2/ERF superfamily has been identified and classified in Hevea brasiliensis. This superfamily includes the ERFs involved in response to ethylene. The relative transcript abundance of ethylene biosynthesis genes and of AP2/ERF genes was analysed during somatic embryogenesis for callus lines with different regeneration potential, in order to identify genes regulated during that process.ResultsThe analysis of relative transcript abundance was carried out by real-time RT-PCR for 142 genes. The transcripts of ERFs from group I, VII and VIII were abundant at all stages of the somatic embryogenesis process. Forty genetic expression markers for callus regeneration capacity were identified. Fourteen markers were found for proliferating calli and 35 markers for calli at the end of the embryogenesis induction phase. Sixteen markers discriminated between normal and abnormal embryos and, lastly, there were 36 markers of conversion into plantlets. A phylogenetic analysis comparing the sequences of the AP2 domains of Hevea and Arabidopsis genes enabled us to predict the function of 13 expression marker genes.ConclusionsThis first characterization of the AP2/ERF superfamily in Hevea revealed dramatic regulation of the expression of AP2/ERF genes during the somatic embryogenesis process. The gene expression markers of proliferating callus capacity to regenerate plants by somatic embryogenesis should make it possible to predict callus lines suitable to be used for multiplication. Further functional characterization of these markers opens up prospects for discovering specific AP2/ERF functions in the Hevea species for which somatic embryogenesis is difficult.

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“…Integumen digunakan sebagai sumber eksplan karena jaringan tersebut paling responsif terhadap kultur in vitro (Montoro, komunikasi pribadi), secara genetik identik dengan tanaman induk, tersusun oleh banyak sel parenkim, dan tervaskularisasi (Carron et al, 1995). Lardet et al (2008) juga mengatakan bahwa eksplan yang mempunyai kompetensi embriogenesis somatik tinggi pada umumnya berasal dari organ reproduktif seperti anter, integumen biji, dan embrio zigotik Media kultur untuk inisasi . kalus adalah MH (Medium for Hevea) yang dikembangkan oleh Carron et al (1989).…”

Section: Inisiasi Kalusunclassified

PERKEMBANGAN KULTUR IN VITRO PADA TANAMAN KARET (Hevea brasiliensis, Müell. Arg.) MELALUI EMBRIOGENESIS SOMATIK DI CIRAD PERANCIS

Bintarti¹

2015

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AbstrakEmbriogenesis somatik menjadi alternatif perbanyakan klonal pada tanaman karet untuk mendapatkan bahan tanam dengan sistem perakaran sendiri dan identik dengan induknya. Penelitian mengenai embriogenesis somatik pada tanaman karet sudah banyak dilakukan oleh lembaga penelitian di dunia, salah satunya adalah lembaga penelitian swasta CIRAD Perancis. Berbagai penelitian dalam upaya pengembangan metode embriogenesis somatik tanaman karet telah diteliti mulai tahun 1970-an, meliputi E m b r i o g e n e s i s S o m a t i k P r i m e r , Embriogenesis Somatik Jangka Panjang atau, dan Embriogenesis Somatik Sekunder. Selain itu, tim peneliti di CIRAD juga telah berhasil mengembangkan teknologi kriopreservasi atau penyimpanan jangka panjang di dalam nitrogen cair yang dikaitkan dengan Embriogenesis Somatic Sekunder untuk menyeleksi dan menetapkan galur-galur kalus yang mempunyai kapasitas embriogenik dan regenerasi tinggi. Walaupun begitu, teknik e m b r i o g e n e s i s s o m a t i k b e l u m b i s a diaplikasikan secara luas untuk perbanyakan tanaman karet secara massal, karena beberapa kendala. Lebih jauh, teknik embriogenesis somatik dan kriopreservasi dapat mendukung p r o g r a m p e mu l i a a n t a n a m a n u n t u k menciptakan tanaman karet transgenik dengan sifat-sifat agronomis unggul seperti toleran terhadap stress abiotik dan produksi lateks tinggi.

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Influences of aging and cloning methods on the capacity for somatic embryogenesis of a mature Hevea brasiliensis genotype

Cited by 31 publications

References 13 publications

Identification of the Hevea brasiliensisAP2/ERF superfamily by RNA sequencing

Identification of the Hevea brasiliensisAP2/ERF superfamily by RNA sequencing

Some ethylene biosynthesis and AP2/ERF genes reveal a specific pattern of expression during somatic embryogenesis in Hevea brasiliensis

PERKEMBANGAN KULTUR IN VITRO PADA TANAMAN KARET (Hevea brasiliensis, Müell. Arg.) MELALUI EMBRIOGENESIS SOMATIK DI CIRAD PERANCIS

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