Characterization and Mutational Analysis of a Monogalactosyldiacylglycerol Synthase Gene OsMGD2 in Rice

Basnet, Rasbin; Zhang, Jiarun; Hussain, N.; Shu, Qingyao

doi:10.3389/fpls.2019.00992

Cited by 25 publications

(30 citation statements)

References 62 publications

(86 reference statements)

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“…Similar results have been found in previous studies, like OsMGD2 and OsDGD2 in rice, which are all localized to chloroplast [75,76]. Their gene knockout plants have reduced fertility.…”

Section: Pigment-related Genes Are Associated With the Phenotype Of Mtsupporting

confidence: 91%

“…Their gene knockout plants have reduced fertility. Additionally, downregulation of OsMGD2 also caused a decrease of chlorophyll content and plant height [76]. PUR4, another chloroplast localized synthase which involved in the purine biosynthetic pathway, is essential to the male gametophyte development [77].…”

Section: Pigment-related Genes Are Associated With the Phenotype Of Mtmentioning

confidence: 99%

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Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Zhang

Liu

et al. 2020

Preprint

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Background: Male sterility is a simple and efficient pollination control system that is widely exploited in hybrid breeding. In upland cotton, CCRI9106, a photosensitive genetic male sterile (PGMS) mutant isolated from CCRI040029, was reported of great advantages to cotton heterosis. However, little information concerning the male sterility of CCRI9106 is known. Here, comparative transcriptome analysis of CCRI9106 (the mutant, MT) and CCRI040029 (the wild type, WT) anthers in Anyang (long-day, male sterile condition to CCRI9106) was performed to reveal the potential male sterile mechanism of CCRI9106.Results: Light and electron microscopy revealed that the male sterility phenotype of MT was mainly attributed to irregularly exine, lacking tryphine and immature anther cuticle. Based on the cytological characteristics of MT anthers, anther RNA libraries (18 in total) of tetrad (TTP), late uninucleate (LUNP) and binucleate (BNP) stages in MT and WT were constructed for transcriptomic analysis, therefore revealing a total of 870,4 differentially expressed genes (DEGs). By performing gene expression pattern analysis and protein-protein interaction (PPI) networks construction, we found down-regulation of DEGs, which enriched by the lipid biosynthetic process and the synthesis pathways of several types of secondary metabolites such as terpenoids, flavonoids and steroids, may crucial to the male sterility phenotype of MT, and resulting in the defects of anther cuticle and tryphine, even the irregularly exine. Furthermore, several lipid-related genes together with ABA-related genes and MYB transcription factors were identified as hub genes via weighted gene co-expression network analysis (WGCNA). Additionally, the ABA content of MT anthers was reduced across all stages when compared with WT anthers. At last, genes related to the formation of anther cuticle and tryphine could activated in MT under short-day condition.Conclusions: We propose that the down-regulation of genes related to the assembly of anther cuticle and tryphine may lead to the male sterile phenotype of MT, and MYB transcription factors together with ABA played key regulatory roles in these processes. The conversion of fertility in different photoperiods may closely relate to the functional expression of these genes. These findings contribute to elucidate the mechanism of male sterility in upland cotton.

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“…Similar results have been found in previous studies, like OsMGD2 and OsDGD2 in rice, which are all localized to chloroplast [75,76]. Their gene knockout plants have reduced fertility.…”

Section: Pigment-related Genes Are Associated With the Phenotype Of Mtsupporting

confidence: 91%

Section: Pigment-related Genes Are Associated With the Phenotype Of Mtmentioning

confidence: 99%

Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Zhang

Liu

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Their gene knockout plants have reduced fertility. Additionally, down-regulation of OsMGD2 also caused a decrease of chlorophyll content and plant height [ 76 ]. PUR4, another chloroplast localized synthase which involved in the purine biosynthetic pathway, is essential to the male gametophyte development [ 77 ].…”

Section: Discussionmentioning

confidence: 99%

Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Zhang

Liu

et al. 2020

BMC Genomics

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Background Male sterility is a simple and efficient pollination control system that is widely exploited in hybrid breeding. In upland cotton, CCRI9106, a photosensitive genetic male sterile (PGMS) mutant isolated from CCRI040029, was reported of great advantages to cotton heterosis. However, little information concerning the male sterility of CCRI9106 is known. Here, comparative transcriptome analysis of CCRI9106 (the mutant, MT) and CCRI040029 (the wild type, WT) anthers in Anyang (long-day, male sterile condition to CCRI9106) was performed to reveal the potential male sterile mechanism of CCRI9106. Results Light and electron microscopy revealed that the male sterility phenotype of MT was mainly attributed to irregularly exine, lacking tryphine and immature anther cuticle. Based on the cytological characteristics of MT anthers, anther RNA libraries (18 in total) of tetrad (TTP), late uninucleate (lUNP) and binucleate (BNP) stages in MT and WT were constructed for transcriptomic analysis, therefore revealing a total of 870,4 differentially expressed genes (DEGs). By performing gene expression pattern analysis and protein-protein interaction (PPI) networks construction, we found down-regulation of DEGs, which enriched by the lipid biosynthetic process and the synthesis pathways of several types of secondary metabolites such as terpenoids, flavonoids and steroids, may crucial to the male sterility phenotype of MT, and resulting in the defects of anther cuticle and tryphine, even the irregularly exine. Furthermore, several lipid-related genes together with ABA-related genes and MYB transcription factors were identified as hub genes via weighted gene co-expression network analysis (WGCNA). Additionally, the ABA content of MT anthers was reduced across all stages when compared with WT anthers. At last, genes related to the formation of anther cuticle and tryphine could activated in MT under short-day condition. Conclusions We propose that the down-regulation of genes related to the assembly of anther cuticle and tryphine may lead to the male sterile phenotype of MT, and MYB transcription factors together with ABA played key regulatory roles in these processes. The conversion of fertility in different photoperiods may closely relate to the functional expression of these genes. These findings contribute to elucidate the mechanism of male sterility in upland cotton.

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“…Genome editing for multiple susceptible genes have been targeted to confer broad spectrum resistance against blight disease in rice [55][56][57]. Subsequently, CRISPR/Cas9 was applied for improving important quantitative and qualitative traits in rice including editing of OsSAPK2 regulating drought responses [58], OsGIF1 regulating size of stem, leaves and grain [59], OsMGD2 regulating grain quality [60], OsARM1 controlling tolerance against for arsenic toxicity [61], OsPPa6 contributing for tolerance against alkalinity [62] and OsBADH2 leading to aroma in rice [35]. Based on the above evidences, this study was undertaken with an aim of demonstrating the efficacy of CRISPR/Cas9 in accelerated development of aromatic rice genotypes and secondly to create different mutations in OsBADH2 leading to accumulation of aroma in rice.…”

Section: Plos Onementioning

confidence: 99%

Creation of novel alleles of fragrance gene OsBADH2 in rice through CRISPR/Cas9 mediated gene editing

et al. 2020

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Fragrance in rice grains is a key quality trait determining its acceptability and marketability. Intensive research on rice aroma identified mutations in betaine aldehyde dehydrogenase (OsBADH2) leading to production of aroma in rice. Gene editing technologies like CRISPR/ Cas9 system has opened new avenues for accelerated improvement of rice grain quality through targeted mutagenesis. In this study, we have employed CRISPR/Cas9 tool to create novel alleles of OsBADH2 leading to introduction of aroma into an elite non-aromatic rice variety ASD16. PCR analysis of putative transformants using primers targeting the flanking regions of sgRNA in the 7 th exon of OsBADH2 identified 37.5% potential multi-allelic mutations in T 0 generation. Sensory evaluation test in the leaves of T 0 lines identified thirteen lines belonging to five independent events producing aroma. Sequence analysis of these aromatic T 0 lines identified 22 different types of mutations located within-17 bp to +15bp of sgRNA region. The-1/-2 bp deletion in the line # 8-19 and-8/-5 bp deletion in the line # 2-16 produced strong aroma and the phenotype was stably inherited in the T 1 generation. Comparative volatile profiling detected novel aromatic compounds viz., pyrrolidine, pyridine, pyrazine, pyradazine and pyrozole in the grains of T 1 progenies of line # 8-19. This study has demonstrated the use of CRISPR/Cas9 in creating novel alleles of OsBADH2 to introduce aroma into any non-aromatic rice varieties.

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Characterization and Mutational Analysis of a Monogalactosyldiacylglycerol Synthase Gene OsMGD2 in Rice

Cited by 25 publications

References 62 publications

Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Deficiencies in the formation and regulation of anther cuticle and tryphine contribute to male sterility in cotton PGMS line

Creation of novel alleles of fragrance gene OsBADH2 in rice through CRISPR/Cas9 mediated gene editing

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