Maize VKS1 Regulates Mitosis and Cytokinesis During Early Endosperm Development

Huang, Yongcai; Wang, Haihai; Huang, Xing; Wang, Qiong; Wang, Jiechen; An, Dong; Li, Jiqin; Wang, Wen-Qin; Wu, Yongrui

doi:10.1105/tpc.18.00966

Cited by 41 publications

(35 citation statements)

References 75 publications

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“…The chromosome separation error in dek15 results in reduced endosperm size and embryo lethality (He et al, 2019). The varied kernel size1 mutant contains a mutation in Zmkin11 that causes impaired spindle assembly, sister chromatid separation, and phragmoplast formation, leading to varying degrees of defective cell proliferation, ultimately resulting in the formation of seeds of different sizes (Huang et al, 2019).…”

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confidence: 99%

Loss of Function of an RNA Polymerase III Subunit Leads to Impaired Maize Kernel Development

et al. 2020

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Kernel size is an important factor determining grain yield. Although a number of genes affecting kernel development in maize (Zea mays) have been identified by analyzing kernel mutants, most of the corresponding mutants cannot be used in maize breeding programs due to low germination or incomplete seed development. Here, we characterized small kernel7, a recessive small-kernel mutant with a mutation in the gene encoding the second-largest subunit of RNA polymerase III (RNAPΙΙΙ; NRPC2). A frame shift in ZmNRPC2 leads to a premature stop codon, resulting in significantly reduced levels of transfer RNAs and 5S ribosomal RNA, which are transcribed by RNAPΙΙΙ. Loss-of-function nrpc2 mutants created by CRISPR/CAS9 showed significantly reduced kernel size due to altered endosperm cell size and number. ZmNRPC2 affects RNAPIII activity and the expression of genes involved in cell proliferation and endoreduplication to control kernel development via physically interacting with RNAPIII subunits RPC53 and AC40, transcription factor class C1 and Floury3. Notably, unlike the semidominant negative mutant floury3, which has defects in starchy endosperm, small kernel7 only affects kernel size but not the composition of kernel storage proteins. Our findings provide novel insights into the molecular network underlying maize kernel size, which could facilitate the genetic improvement of maize in the future.

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Loss of Function of an RNA Polymerase III Subunit Leads to Impaired Maize Kernel Development

et al. 2020

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“…Recently, advanced genomic and molecular tools have been applied to characterize a wide-array of specific genes’ functions associated with the ESD, such as the regulation of endosperm cell proliferation [ 6 , 7 ] and accumulation of storage components [ 8 – 11 ]. Complex interactions exist among the key regulators and related genes involved in these processes.…”

Section: Introductionmentioning

confidence: 99%

Optimization of isolation and transfection conditions of maize endosperm protoplasts

Song

Gao

et al. 2020

Plant Methods

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Background Endosperm-trait related genes are associated with grain yield or quality in maize. There are vast numbers of these genes whose functions and regulations are still unknown. The biolistic system, which is often used for transient gene expression, is expensive and involves complex protocol. Besides, it cannot be used for simultaneous analysis of multiple genes. Moreover, the biolistic system has little physiological relevance when compared to cell-specific based system. Plant protoplasts are efficient cell-based systems which allow quick and simultaneous transient analysis of multiple genes. Typically, PEG-calcium mediated transfection of protoplast is simple and cost-effective. Notably, starch granules in cereal endosperm may diminish protoplast yield and integrity, if the isolation and transfection conditions are not accurately measured. Prior to this study, no PEG-calcium mediated endosperm protoplast system has been reported for cereal crop, perhaps, because endosperm cells accumulate starch grains. Results Here, we showed the uniqueness of maize endosperm-protoplast system (EPS) in conducting endosperm cell-based experiments. By using response surface designs, we established optimized conditions for the isolation and PEG-calcium mediated transfection of maize endosperm protoplasts. The optimized conditions of 1% cellulase, 0.75% macerozyme and 0.4 M mannitol enzymolysis solution for 6 h showed that more than 80% protoplasts remained viable after re-suspension in 1 ml MMG. The EPS was used to express GFP protein, analyze the subcellular location of ZmBT1, characterize the interaction of O2 and PBF1 by bimolecular fluorescent complementation (BiFC), and simultaneously analyze the regulation of ZmBt1 expression by ZmMYB14. Conclusions The described optimized conditions proved efficient for reasonable yield of viable protoplasts from maize endosperm, and utility of the protoplast in rapid analysis of endosperm-trait related genes. The development of the optimized protoplast isolation and transfection conditions, allow the exploitation of the functional advantages of protoplast system over biolistic system in conducting endosperm-based studies (particularly, in transient analysis of genes and gene regulation networks, associated with the accumulation of endosperm storage products). Such analyses will be invaluable in characterizing endosperm-trait related genes whose functions have not been identified. Thus, the EPS will benefit the research of cereal grain yield and quality improvement.

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“…Diversity in endosperm ploidy level of angiosperms broadly ranges from 2n to 15n, especially due to the variability in the genetic contribution of the female parent from 1n to 14n (Friedman et al, 2008). Key genes regulating the cell cycle, mega-sporogenesis or -gametogenesis or endosperm developmental process, including ontogeny and cell numbers are well documented (Motamayor et al, 2000; Berger et al, 2006; Huh et al, 2007; Fiume and Fletcher, 2012; Batista et al, 2019; Huang et al, 2019; Kirkbride et al, 2019). However, the variability underlying these traits between species and its regulatory mechanism that flexibly accommodates and accepts the variable endosperm dosage level from 2n to 15n, and endosperm cell number in angiosperms is not yet fully understood.…”

Section: Introductionmentioning

confidence: 99%

Endosperm variability: from endoreduplication within a seed to higher ploidy across species, and its competence

Rangan

2020

Seed Sci. Res.

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Endosperm tissue that nourishes the embryo during seed development, upon maturity, nourishes the global population with special reference to cereal crops like maize, wheat and rice. In about 70% of the angiosperms, endosperm genome content is ‘3n’ with 2:1 (maternal:paternal) contribution, as a result of the second fertilization event. However, angiosperms evolution also documents diversity in endosperm genome content from ‘2n’ to ‘15n’, in scale with the corresponding maternal genome dosage variability (‘1n’ to ‘14n’), whereas paternal contribution is invariable. In apomicts, due to lack of fertilization, or pseudogamy (fertilization of the central cell for endosperm formation), endosperm genome dosage (m:p) has been reported to range between 1:1 and 8:3. Exceptionally, the central cell with one unreduced nucleus and fused with a reduced sperm cell, with 2:1 normal genome dosage, has been reported in Panicum. Altered genome dosage levels are reportedly correlative with eccentricities among maternal and paternal contribution to seed resource allocation. Besides endosperm ploidy variability between species of angiosperms, the present review gives an overview of the ploidy variability in endosperm cells within a seed, up to ‘690n’. In addition to genome-scale variability in the endosperm, some taxa of angiosperms exhibit chlorophyllous endosperms and some chlorophyllous embryos. Also, endosperm cell number during seed development is reported to have a strong association with grain weight at maturity. Genes underlying these traits of variability are unknown, and the present review underscores the variability and highlights the potential of the single-cell sequencing techniques towards understanding the genetic mechanisms associated with these variable traits.

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Maize VKS1 Regulates Mitosis and Cytokinesis During Early Endosperm Development

Cited by 41 publications

References 75 publications

Loss of Function of an RNA Polymerase III Subunit Leads to Impaired Maize Kernel Development

Loss of Function of an RNA Polymerase III Subunit Leads to Impaired Maize Kernel Development

Optimization of isolation and transfection conditions of maize endosperm protoplasts

Endosperm variability: from endoreduplication within a seed to higher ploidy across species, and its competence

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