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AtWuschel promotes formation of the embryogenic callus in Gossypium hirsutum.

ABSTRACT: Upland cotton (Gossypium hirsutum) is one of the most recalcitrant species for in vitro plant regeneration through somatic embryogenesis. Callus from only a few cultivars can produce embryogenic callus (EC), but the mechanism is not well elucidated. Here we screened a cultivar, CRI24, with high efficiency of EC produce. The expression of genes relevant to EC production was analyzed between the materials easy to or difficult to produce EC. Quantitative PCR showed that CRI24, which had a 100% EC differentiation rate, had the highest expression of the genes GhLEC1, GhLEC2, and GhFUS3. Three other cultivars, CRI12, CRI41, and Lu28 that formed few ECs expressed these genes only at low levels. Each of the genes involved in auxin transport (GhPIN7) and signaling (GhSHY2) was most highly expressed in CRI24, with low levels in the other three cultivars. WUSCHEL (WUS) is a homeodomain transcription factor that promotes the vegetative-to-embryogenic transition. We thus obtained the calli that ectopically expressed Arabidopsis thaliana Wus (AtWus) in G. hirsutum cultivar CRI12, with a consequent increase of 47.75% in EC differentiation rate compared with 0.61% for the control. Ectopic expression of AtWus in CRI12 resulted in upregulation of GhPIN7, GhSHY2, GhLEC1, GhLEC2, and GhFUS3. AtWus may therefore increase the differentiation potential of cotton callus by triggering the auxin transport and signaling pathways.

SUBMITTER: Zheng W

PROVIDER: S-EPMC3909107 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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AtWuschel promotes formation of the embryogenic callus in Gossypium hirsutum.

Zheng Wu W Zhang Xueyan X Yang Zuoren Z Wu Jiahe J Li Fenglian F Duan Lanling L Liu Chuanliang C Lu Lili L Zhang Chaojun C Li Fuguang F

PloS one 20140131 1

Upland cotton (Gossypium hirsutum) is one of the most recalcitrant species for in vitro plant regeneration through somatic embryogenesis. Callus from only a few cultivars can produce embryogenic callus (EC), but the mechanism is not well elucidated. Here we screened a cultivar, CRI24, with high efficiency of EC produce. The expression of genes relevant to EC production was analyzed between the materials easy to or difficult to produce EC. Quantitative PCR showed that CRI24, which had a 100% EC d ...[more]

PMID: 24498119

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Project description:BackgroundBarley is known to be recalcitrant to tissue culture, which hinders genetic transformation and its biotechnological application. To date, the ideal explant for transformation remains limited to immature embryos; the mechanism underlying embryonic callus formation is elusive.ResultsThis study aimed to uncover the different transcription regulation pathways between calli formed from immature (IME) and mature (ME) embryos through transcriptome sequencing. We showed that incubation of embryos in an auxin-rich medium caused dramatic changes in gene expression profiles within 48 h. Overall, 9330 and 11,318 differentially expressed genes (DEGs) were found in the IME and ME systems, respectively. 3880 DEGs were found to be specific to IME_0h/IME_48h, and protein phosphorylation, regulation of transcription, and oxidative-reduction processes were the most common gene ontology categories of this group. Twenty-three IAA, fourteen ARF, eight SAUR, three YUC, and four PIN genes were found to be differentially expressed during callus formation. The effect of callus-inducing medium (CIM) on IAA genes was broader in the IME system than in the ME system, indicating that auxin response participates in regulating cell reprogramming during callus formation. BBM, LEC1, and PLT2 exhibited a significant increase in expression levels in the IME system but were not activated in the ME system. WUS showed a more substantial growth trend in the IME system than in the ME system, suggesting that these embryonic, shoot, and root meristem genes play crucial roles in determining the acquisition of competency. Moreover, epigenetic regulators, including SUVH3A, SUVH2A, and HDA19B/703, exhibited differential expression patterns between the two induction systems, indicating that epigenetic reprogramming might contribute to gene expression activation/suppression in this process. Furthermore, we examined the effect of ectopic expression of HvBBM and HvWUS on Agrobacterium-mediated barley transformation. The transformation efficiency in the group expressing the PLTPpro:HvBBM + Axig1pro:HvWUS construct was increased by three times that in the control (empty vector) because of enhanced plant regeneration capacity.ConclusionsWe identified some regulatory factors that might contribute to the differential responses of the two explants to callus induction and provide a promising strategy to improve transformation efficiency in barley.

Dataset Information

AtWuschel promotes formation of the embryogenic callus in Gossypium hirsutum.

Publications

AtWuschel promotes formation of the embryogenic callus in Gossypium hirsutum.

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