Leaf development has been extensively studied on a genetic level. However, little is known about the inter- play between the developmental regulators and the cell cycle machinery--a link that ultimately affects leaf f...Leaf development has been extensively studied on a genetic level. However, little is known about the inter- play between the developmental regulators and the cell cycle machinery--a link that ultimately affects leaf form and size. miR319 is a conserved microRNA that regulates TCP transcription factors involved in multiple developmental pathways, including leaf development and senescence, organ curvature, and hormone biosynthesis and signaling. Here, we analyze the participation of TCP4 in the control of cell proliferation. A small increase in TCP4 activity has an immediate impact on leaf cell number, by significantly reducing cell proliferation. Plants with high TCP4 levels have a strong reduction in the expression of genes known to be active in G2-M phase of the cell cycle. Part of these effects is mediated by induction of miR396, which represses Growth-Regulating Factor （GRF） transcription factors. Detailed analysis revealed TCP4 to be a direct regulator of MIR396b. However, we found that TCP4 can control cell proliferation through additional pathways, and we identified a direct connection between TCP4 and ICK1/KRP1, a gene involved in the progression of the cell cycle. Our results show that TCP4 can activate different pathways that repress cell proliferation.展开更多
Yield improvement has long been an important task for soybean breeding in the world in order to meet the increasing demand for food and animal feed.mi R396 genes have been shown to negatively regulate grain size in ri...Yield improvement has long been an important task for soybean breeding in the world in order to meet the increasing demand for food and animal feed.mi R396 genes have been shown to negatively regulate grain size in rice,but whether mi R396 family members may function in a similar manner in soybean is unknown.Here,we generated eight soybean mutants harboring different combinations of homozygous mutations in the six soybean mi R396genes through genome editing with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated nuclease(Cas)12SF01 in the elite soybean cultivar Zhonghuang 302(ZH302).Four triple mutants(mir396aci,mir396acd,mir396adf,and mir396cdf),two quadruple mutants(mir396-abcd and mir396acfi),and two quintuple mutants(mir396abcdf and mir396bcdfi)were characterized.We found that plants of all the mir396 mutants produced larger seeds compared to ZH302 plants.Field tests showed that mir396adf and mir396cdf plants have significantly increased yield in growth zones with relatively high latitude which are suited for ZH302 and moderately increased yield in lower latitude.In contrast,mir396abcdf and mir396bcdfiplants have increased plant height and decreased yield in growth zones with relatively high latitude due to lodging issues,but they are suited for low latitude growth zones with increased yield without lodging problems.Taken together,our study demonstrated that loss-of-function of mi R396 genes leads to significantly enlarged seed size and increased yield in soybean,providing valuable germplasms for breeding high-yield soybean.展开更多
Secondary vascular development is a key biological characteristic of woody plants and the basis of wood formation.Our understanding of gene expression regulation and dynamic changes in microRNAs(miRNAs)during secondar...Secondary vascular development is a key biological characteristic of woody plants and the basis of wood formation.Our understanding of gene expression regulation and dynamic changes in microRNAs(miRNAs)during secondary vascular development is still limited.Here we present an integrated analysis of the miRNA and mRNA transcriptome of six phase-specific tissues-the shoot apex,procambium,primary vascular tissue,cambium,secondary phloem,and secondary xylem-in Populus tomentosa.Several novel regulatory modules,including the PtoTCP20-miR396d-PtoGRF15 module,were identified during secondary vascular development in Populus.A series of biochemical and molecular experiments confirmed that PtoTCP20 activated transcription of the miR396d precursor gene and that miR396d targeted PtoGRF15 to downregulate its expression.Plants overexpressing miR396d(35S:miR396d)showed enhanced secondary growth and increased xylem production.Conversely,during the transition from primary to secondary vascular development,plants with downregulated PtoTCP20expression(PtoTCP20-SRDX),downregulated miR396 expression(35S:STTM396),and PtoGRF15 overexpression(35S:PtoGRF15)showed delayed secondary growth.Novel regulatory modules were identified by integrated analysis of the miRNA and mRNA transcriptome,and the regulatory role of the PtoTCP20-miR396d-PtoGRF15 signaling cascade in secondary vascular development was validated in Populus,providing information to support improvements in forest cultivation and wood properties.展开更多
Grain size determines grain weight and affects grain quality. Several major quantitative trait loci （QTLs） regulating grain size have been cloned; however, our understanding of the underlying mechanism that regulate...Grain size determines grain weight and affects grain quality. Several major quantitative trait loci （QTLs） regulating grain size have been cloned; however, our understanding of the underlying mechanism that regulates the size of rice grains remains fragmentary. Here, we report the cloning and characterization of a dominant QTL, GRAIN SIZE ON CHROMOSOME 2 （GS2）, which encodes Growth-Regulating Factor 4 （OsGRF4）, a transcriptional regulator. GS2 localizes to the nucleus and may act as a transcription activator. A rare mutation of GS2 affecting the binding site of a microRNA, OsmiR396c, causes elevated expression of GS2/OsGRF4. The increase in GS2 expression leads to larger cells and increased numbers of cells, which thus enhances grain weight and yield. The introduction of this rare allele of GS2/OsGRF4 into rice cultivars could significantly enhance grain weight and increase grain yield, with possible applications in breeding high-yield rice varieties.展开更多
文摘Leaf development has been extensively studied on a genetic level. However, little is known about the inter- play between the developmental regulators and the cell cycle machinery--a link that ultimately affects leaf form and size. miR319 is a conserved microRNA that regulates TCP transcription factors involved in multiple developmental pathways, including leaf development and senescence, organ curvature, and hormone biosynthesis and signaling. Here, we analyze the participation of TCP4 in the control of cell proliferation. A small increase in TCP4 activity has an immediate impact on leaf cell number, by significantly reducing cell proliferation. Plants with high TCP4 levels have a strong reduction in the expression of genes known to be active in G2-M phase of the cell cycle. Part of these effects is mediated by induction of miR396, which represses Growth-Regulating Factor （GRF） transcription factors. Detailed analysis revealed TCP4 to be a direct regulator of MIR396b. However, we found that TCP4 can control cell proliferation through additional pathways, and we identified a direct connection between TCP4 and ICK1/KRP1, a gene involved in the progression of the cell cycle. Our results show that TCP4 can activate different pathways that repress cell proliferation.
基金National High Technology Research and Development Program of China(863 Program)(2006AA02Z129)National Natural Science Foundation of China(90408022)+1 种基金Science Foundation of Yunnan Province(2004C0051M)"Hundredtalents" Program of the Chinese Academy of Sciences
基金supported by Bel agen Biotechnology Inc.,and by the National Key Research and Development Program of China(No.NK2022010301)the Seed-Industrialized Development Program in Shandong Province,China(No.2021LZGC003)+3 种基金the Key Research&Development Program of Shandong Province,China(No.2021LZGC012)the Innovative research team of Anhui Education(No.2022AH010056)the National Natural Science Foundation of China(No.32270367)the Ph D Scientific Research and Innovation Foundation of Sanya Yazhou Bay Science and Technology City(No.HSPHDSRF-2023-11-007)。
文摘Yield improvement has long been an important task for soybean breeding in the world in order to meet the increasing demand for food and animal feed.mi R396 genes have been shown to negatively regulate grain size in rice,but whether mi R396 family members may function in a similar manner in soybean is unknown.Here,we generated eight soybean mutants harboring different combinations of homozygous mutations in the six soybean mi R396genes through genome editing with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated nuclease(Cas)12SF01 in the elite soybean cultivar Zhonghuang 302(ZH302).Four triple mutants(mir396aci,mir396acd,mir396adf,and mir396cdf),two quadruple mutants(mir396-abcd and mir396acfi),and two quintuple mutants(mir396abcdf and mir396bcdfi)were characterized.We found that plants of all the mir396 mutants produced larger seeds compared to ZH302 plants.Field tests showed that mir396adf and mir396cdf plants have significantly increased yield in growth zones with relatively high latitude which are suited for ZH302 and moderately increased yield in lower latitude.In contrast,mir396abcdf and mir396bcdfiplants have increased plant height and decreased yield in growth zones with relatively high latitude due to lodging issues,but they are suited for low latitude growth zones with increased yield without lodging problems.Taken together,our study demonstrated that loss-of-function of mi R396 genes leads to significantly enlarged seed size and increased yield in soybean,providing valuable germplasms for breeding high-yield soybean.
基金This work was supported by the National Natural Science Foundation of China(32071726 and 32271825).
文摘Secondary vascular development is a key biological characteristic of woody plants and the basis of wood formation.Our understanding of gene expression regulation and dynamic changes in microRNAs(miRNAs)during secondary vascular development is still limited.Here we present an integrated analysis of the miRNA and mRNA transcriptome of six phase-specific tissues-the shoot apex,procambium,primary vascular tissue,cambium,secondary phloem,and secondary xylem-in Populus tomentosa.Several novel regulatory modules,including the PtoTCP20-miR396d-PtoGRF15 module,were identified during secondary vascular development in Populus.A series of biochemical and molecular experiments confirmed that PtoTCP20 activated transcription of the miR396d precursor gene and that miR396d targeted PtoGRF15 to downregulate its expression.Plants overexpressing miR396d(35S:miR396d)showed enhanced secondary growth and increased xylem production.Conversely,during the transition from primary to secondary vascular development,plants with downregulated PtoTCP20expression(PtoTCP20-SRDX),downregulated miR396 expression(35S:STTM396),and PtoGRF15 overexpression(35S:PtoGRF15)showed delayed secondary growth.Novel regulatory modules were identified by integrated analysis of the miRNA and mRNA transcriptome,and the regulatory role of the PtoTCP20-miR396d-PtoGRF15 signaling cascade in secondary vascular development was validated in Populus,providing information to support improvements in forest cultivation and wood properties.
文摘Grain size determines grain weight and affects grain quality. Several major quantitative trait loci （QTLs） regulating grain size have been cloned; however, our understanding of the underlying mechanism that regulates the size of rice grains remains fragmentary. Here, we report the cloning and characterization of a dominant QTL, GRAIN SIZE ON CHROMOSOME 2 （GS2）, which encodes Growth-Regulating Factor 4 （OsGRF4）, a transcriptional regulator. GS2 localizes to the nucleus and may act as a transcription activator. A rare mutation of GS2 affecting the binding site of a microRNA, OsmiR396c, causes elevated expression of GS2/OsGRF4. The increase in GS2 expression leads to larger cells and increased numbers of cells, which thus enhances grain weight and yield. The introduction of this rare allele of GS2/OsGRF4 into rice cultivars could significantly enhance grain weight and increase grain yield, with possible applications in breeding high-yield rice varieties.
