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Establishment of an Efficient Regeneration System in Goodyera foliosa and Comprehensive Analysis of Functionally Regulated Genes Involved in Developmental Regulatory Pathways Based on Transcriptome Analysis
HE Guan-rong,HE Bi-zhu,WU Sha-sha,SHI Jing-shan,CHEN Ji-shuang,LAN Si-ren
China Biotechnology ›› 2018, Vol. 38 ›› Issue (12) : 57-64.
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Establishment of an Efficient Regeneration System in Goodyera foliosa and Comprehensive Analysis of Functionally Regulated Genes Involved in Developmental Regulatory Pathways Based on Transcriptome Analysis
({{custom_author.role_en}}), {{javascript:window.custom_author_en_index++;}}Goodyera foliosa, belonged to the genus of Cymbidium, is an endangered wild and national secondary protected plant which is used as ornamental plants and for various medicinal purposes. Duo to its small distribution population and weak transmission and diffusion, the natural reproduction is greatly limited. In this study, a high efficient in vitro regeneration system was developed from stem explants of Goodyera foliosa. The functional genes involved in the morphogenesis development was deeply explored by integrating with high-throughput transcriptome sequencing and bioinformatics analysis technology. For shoot-inducing , the optical culture medium is Morel + 2.0 mg/L 6-BA + 0.5 mg/L KT+1.0 mg/L NAA + 1g/L peptone + 25g/L sucrose + 7.0 g/L Agar + 1.0 g/L active carbon + 30 g/L banana + 50 g/L potato. The optical culture medium for bud proliferation is Morel + 3 mg/L 6-BA + 0.5 mg/L NAA + 0.5 mg/L KT + 0.01 mg/L TDZ + 2g/L peptone + 25g/L sucrose + 7.0 g/L Agar + 1.0 g/L active carbon + 30g/L banana + 50g/L potato. On the rooting medium with 1/2 Morel + 1.0 mg/L IBA + 0.1 mg/L NAA + 1 g/L + Hyponex NO.2 + 25g/L sucrose + 7.0g/L Agar + 1.0 g/L active carbon + 1g/L peptone. After transcriptom sequencing and assembling, 170, 688 Unigenes were obtained. The average length and N50 length of Unigenes was 584bp and 833bp respectively. Total of 17, 352 Unigenes were completely annotated to 5 functional databases including NR, Swiss-Prot, KOG, GO and KEGG. The functional analysis of differential Unigenes was showed that hormone signal transduction, plant development, secondary metabolites and energy metabolism were significantly enriched. Moreover, 511 Unigene encoding transcription factors involved with plant organ developmental regulation were predicted. Conclusion, a comprehensive transcriptom landscape of Goodyera foliosa was described by integrating with a high efficient in vitro regeneration system and next high-throughput trancriptom sequencing. This work could provide certain reference for fast propagation, genetic transformation, functional gene mining and development mechanism research of Goodyera foliosa.
Goodyera foliosa (Lindl) Benth. / Bioresource conservation / Seedling regeneration Organ development / Transcriptome analysis {{custom_keyword}} /
Table 1 Effect of 6-BA on inducing multiple shoots表1 6-BA含量对芽诱导的影响 |
| 6-BA激素配比 6-BA (mg/L) | 外植体数 Number of explants | 增殖倍数 Proliferation rate | 生长态势 Growth situation |
|---|---|---|---|
| 1.0 | 50 | 2.22±0.03c | 芽少,细弱,生长慢 |
| 1.5 | 50 | 2.61±0.02b | 芽多,壮实,生长块 |
| 2.0 | 50 | 3.21±0.03a | 芽多,壮实,生长块 |
| 2.5 | 50 | 2.11±0.03d | 芽少,壮实,生长慢 |
| 3.0 | 50 | 1.40±0.03e | 芽少,细弱,生长慢 |
Table 2 Effect of NAA on bud multiplication表2 NAA含量对芽增殖的影响 |
| NAA激素配比 NAA (mg/L) | 增殖倍数 Proliferation rate | 生长态势 Growth situation |
|---|---|---|
| 0.1 | 0.92±0.02e | 少量芽、稍绿、长势弱 |
| 0.5 | 1.61±0.02d | 少量芽、稍绿、长势弱 |
| 1.0 | 2.34±0.04b | 芽健壮、浓绿、生长正常 |
| 1.5 | 2.80±0.03a | 芽健壮、浓绿、生长正常 |
| 2.0 | 2.10±0.03c | 芽健壮、绿、生长弱 |
Table 3 Effect of IBA levels on shoot rooting of the plantlet表3 IBA的含量对生根的影响 |
| IBA激素配比 IBA(mg/L) | 株数 Number of seedlings | 生根数 Root number | 平均根长 Mean length of root |
|---|---|---|---|
| 0.5 | 50 | 31.20±1.60b | 1.92±0.02c |
| 1.0 | 50 | 38.23±0.18a | 2.63±0.03a |
| 1.5 | 50 | 29.93±0.18b | 2.10±0.01b |
| 2.0 | 50 | 24.00±0.21c | 1.61±0.02d |
| 2.5 | 50 | 20.13±0.23d | 1.39±0.01e |
Fig.3 GO function analysis of differential expressed Unigenes图3 差异Unigene GO功能富集结果 |
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The authors have declared that no competing interests exist.
作者已声明无竞争性利益关系。
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Table 1 Effect of 6-BA on inducing multiple shootsTable 2 Effect of NAA on bud multiplicationTable 3 Effect of IBA levels on shoot rooting of the plantlet
Fig.1 Length distribution of UnigenesFig.2 Veen diagram of annotated UnigenesFig.3 GO function analysis of differential expressed UnigenesFig.4 KEGG function analysis of differential expressed Unigenes/
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