Gene expression and metabolite profiling of gibberellin biosynthesis during induction of somatic embryogenesis in Medicago truncatula Gaertn

Igielski, Rafał; Kępczyńska, Ewa

doi:10.1371/journal.pone.0182055

Cited by 36 publications

(33 citation statements)

References 108 publications

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“…The GA biosynthesis pathway involves many genes besides CPS ( Figure 5 B). According to the reference [ 54 ], we tested the expression level of the putative genes ( Table S2 ) in the GA biosynthesis pathway of M. truncatula stem tissue. The results showed that most GA biosynthesis genes were highly upregulated in the mnp1-1 mutant, while a small proportion of the genes ( MtKS , MtKAO1 , MtCYP714_A1 , MtCYP714_C2 ) showed a low level of upregulation, with no significant difference.…”

Section: Resultsmentioning

confidence: 99%

Cloning and Functional Analysis of Dwarf Gene Mini Plant 1 (MNP1) in Medicago truncatula

Guo

Zhang

Bai

et al. 2020

IJMS

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Plant height is a vital agronomic trait that greatly determines crop yields because of the close relationship between plant height and lodging resistance. Legumes play a unique role in the worldwide agriculture; however, little attention has been given to the molecular basis of their height. Here, we characterized the first dwarf mutant mini plant 1 (mnp1) of the model legume plant Medicago truncatula. Our study found that both cell length and the cell number of internodes were reduced in a mnp1 mutant. Using the forward genetic screening and subsequent whole-genome resequencing approach, we cloned the MNP1 gene and found that it encodes a putative copalyl diphosphate synthase (CPS) implicated in the first step of gibberellin (GA) biosynthesis. MNP1 was highly homologous to Pisum sativum LS. The subcellular localization showed that MNP1 was located in the chloroplast. Further analysis indicated that GA3 could significantly restore the plant height of mnp1-1, and expression of MNP1 in a cps1 mutant of Arabidopsis partially rescued its mini-plant phenotype, indicating the conservation function of MNP1 in GA biosynthesis. Our results provide valuable information for understanding the genetic regulation of plant height in M. truncatula.

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Section: Resultsmentioning

confidence: 99%

Cloning and Functional Analysis of Dwarf Gene Mini Plant 1 (MNP1) in Medicago truncatula

Guo

Zhang

Bai

et al. 2020

IJMS

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“…The phylogenetic tree was constructed using the Maximum-Likelihood method in MEGA7 41 , with bootstrap analysis of 1,000 replicates for stability testing of the tree nodes. Identification of other GA biosynthesis pathway genes, including CPS , KS , KO , KAO , GA13OX , and GA3ox , in the M. truncatula genome (Mt4.0) was also performed by BLASTP search (E value cutoff of E −10 and query coverage of 50%) using the previously reported GA biosynthesis genes of M. truncatula 42 as the seed queries.…”

Section: Methodsmentioning

confidence: 99%

MtGA2ox10 encoding C20-GA2-oxidase regulates rhizobial infection and nodule development in Medicago truncatula

Kim

Son

et al. 2019

Sci Rep

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Gibberellin (GA) plays a controversial role in the legume-rhizobium symbiosis. Recent studies have shown that the GA level in legumes must be precisely controlled for successful rhizobial infection and nodule organogenesis. However, regulation of the GA level via catabolism in legume roots has not been reported to date. Here, we investigate a novel GA inactivating C20-GA2-oxidase gene MtGA2ox10 in Medicago truncatula . RNA sequencing analysis and quantitative polymerase chain reaction revealed that MtGA2ox10 was induced as early as 6 h post-inoculation (hpi) of rhizobia and reached peak transcript abundance at 12 hpi. Promoter::β-glucuronidase fusion showed that the promoter activity was localized in the root infection/differentiation zone during the early stage of rhizobial infection and in the vascular bundle of the mature nodule. The CRISPR/Cas9-mediated deletion mutation of MtGA2ox10 suppressed infection thread formation, which resulted in reduced development and retarded growth of nodules on the Agrobacterium rhizogenes -transformed roots. Over-expression of MtGA2ox10 in the stable transgenic plants caused dwarfism, which was rescued by GA 3 application, and increased infection thread formation but inhibition of nodule development. We conclude that MtGA2ox10 plays an important role in the rhizobial infection and the development of root nodules through fine catabolic tuning of GA in M. truncatula .

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“…Furthermore, Jiménez and Bangerth ( 2000 , 2001a , b ) and Jiménez et al ( 2005 ) found no difference between GAs levels in cultures of grapevine, carrot, and wheat showing different embryogenic characteristics. Previously, we showed that during a 3-week induction phase in Medicago truncatula, when the cells of the leaf explant from embryogenic (M9-10a) and non-embryogenic (M9) genotypes dedifferentiate to form callus, all the active gibberellins from 13-hydroxylation pathway (GA 1 , GA 3 , GA 6 ) as well as from the non-13-hydroxylation pathway (GA 4 , GA 7 ) are present (Igielski and Kępczyńska 2017 ). However, the contents of GA 3 and also of GA 6 were observed to increase in the M9-10a genotype as the induction phase progressed.…”

Section: Introductionmentioning

confidence: 99%

Profiles of endogenous ABA, bioactive GAs, IAA and their metabolites in Medicago truncatula Gaertn. non-embryogenic and embryogenic tissues during induction phase in relation to somatic embryo formation

Kępczyńska

Orłowska

2021

Planta

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Main conclusion During the 3-week-long induction phase, when M. truncatula cells leaf explants from non-embryogenic genotype (M9) and embryogenic variant (M9-10a) were forming the callus, biosynthesis and degradation of ABA, Gas and IAA proceeded at different levels. Induction of embryo formation is related to a lower ABA content, compared to the content of IAA and that of total bioactive GAs. Abstract Endogenous phytohormones are involved in the regulation of zygotic embryogenesis, but their role, especially of ABA, a plant growth inhibitor, in inducing somatic embryogenesis (SE) in angiosperms is still incompletely known. To arrive a better understanding of the ABA role in the process, we analyzed simultaneously and in detail changes in the contents of both ABA and five bioactive GAs (GA4, GA7, GA1, GA3, GA6) and IAA in M. truncatula non-embryogenic M9 (NE) and embryogenic M9-10a (E) genotypes. The initial leaf explants of both genotypes, and particularly NE, contained many times more ABA compared to the total bioactive GAs or IAA. In tissues during the entire 21-day induction all the hormones mentioned and their metabolites or conjugates were present; however, their contents were found to differ between the lines tested. The ABA level in primary explants of NE genotype was more than two times higher than that in E genotype. An even larger difference in the ABA content was found on the last day (day 21) of the induction phase (IP); the ABA content in E callus was over six times lower than in NE callus. In contrast, the IAA and GAs contents in primary explants of both genotypes in relation to ABA were low, but the contents of IAA and GAs exceeded that of ABA in the M9-10a tissues on the last day of IP. It is shown for the first time that endogenous ABA together with endogenous bioactive GAs and IAA is involved in acquisition of embryogenic competence in Medicago truncatula leaf somatic cells. These findings have a strong functional implication as they allow to improve the SE induction protocol.

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Gene expression and metabolite profiling of gibberellin biosynthesis during induction of somatic embryogenesis in Medicago truncatula Gaertn

Cited by 36 publications

References 108 publications

Cloning and Functional Analysis of Dwarf Gene Mini Plant 1 (MNP1) in Medicago truncatula

Cloning and Functional Analysis of Dwarf Gene Mini Plant 1 (MNP1) in Medicago truncatula

MtGA2ox10 encoding C20-GA2-oxidase regulates rhizobial infection and nodule development in Medicago truncatula

Profiles of endogenous ABA, bioactive GAs, IAA and their metabolites in Medicago truncatula Gaertn. non-embryogenic and embryogenic tissues during induction phase in relation to somatic embryo formation

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