Dek219 encodes the DICER-LIKE1 protein that affects chromatin accessibility and kernel development in maize

Chromatin accessibility plays a vital role in gene transcriptional regulation.However,the regulatory mechanism of chromatin accessibility,as well as its role in regulating crucial gene expression and kernel development in maize(Zea mays)are poorly understood.In this study,we isolated a maize kernel mutant designated as defective kernel219(dek219),which displays opaque endosperm and embryo abortion.Dek219 encodes the DICER-LIKE1(DCL1)protein,an essential enzyme in mi RNA biogenesis.Loss of function of Dek219 results in significant reductions in the expression levels of most mi RNAs and histone genes.Further research showed that the Heat shock transcription factor17(Hsf17)-Zm00001d016571 module may be one of the factors affecting the expression of histone genes.Assay results for transposase-accessible chromatin sequencing(ATAC-seq)indicated that the chromatin accessibility of dek219 is altered compared with that of wild type(WT),which may regulate the expression of crucial genes in kernel development.By analyzing differentially expressed genes(DEGs)and differentially accessible chromatin regions(ACRs)between WT and dek219,we identified 119 candidate genes that are regulated by chromatin accessibility,including some reported to be crucial genes for kernel development.Taken together,these results suggest that Dek219 affects chromatin accessibility and the expression of crucial genes that are required for maize kernel development.

Characterization of transgenic wheat lines expressing maize ABP7 involved in kernel development

Wheat is one of the major food crops in the world.Functional validation of the genes in increasing the grain yield of wheat by genetic engineering is essential for feeding the ever-growing global population.This study investigated the role of ABP7,a bHLH transcription factor from maize involved in kernel development,in regulating grain yield-related traits in transgenic wheat.Molecular characterization showed that transgenic lines HB123 and HB287 contained multicopy integration of ABP7 in the genome with higher transgene expression.At the same time,QB205 was a transgenic event of single copy insertion with no significant difference in ABP7 expression compared to wild-type(WT) plants.Phenotyping under field conditions showed that ABP7 over-expressing transgenic lines HB123 and HB287 exhibited improved grain yield-related traits(e.g.,grain number per spike,grain weight per spike,thousand-grain weight,grain length,and grain width) and increased grain yield per plot,compared to WT plants,whereas line QB205 did not.In addition,total chlorophyll,chlorophyll a,chlorophyll b,and total soluble sugars were largely increased in the flag leaves of both HB123and HB287 transgenic lines compared to the WT.These results strongly suggest that ABP7 positively regulates yieldrelated traits and plot grain yield in transgenic wheat.Consequently,ABP7 can be utilized in wheat breeding for grain yield improvement.

Maize orthologs of rice GS5 and their transregulator are associated with kernel development

Genome information from model species such as rice can assist in the cloning of genes in a complex genome, such as maize. Here, we identified a maize ortholog of rice GS5 that contributes to kernel development in maize. The genome- wide association analysis of the expression levels of ZmGSs, and 15 of its 26 paralogs, identified a trans-regulator on chromosome 7, which was a BAK1-1ike gene. This gene that we named as ZmBAK1-7 could regulate the expression of ZmGS5 and three of the paralogs. Candidate-gene association analyses revealed that these five genes were associated with maize kernel development-related traits. Linkage analyses also detected that ZINGS5 and ZmI3AK1-7 co-localized with mapped QTLs. A transgenic analysis of ZINGS5 in Arabidopsis thaliana L. showed a significant increase in seed weight and cell number, suggesting that 2mG55 may have a conserved function among different plant species that affects seed development.

早实核桃种仁油脂积累过程中主要营养物质的变化

【目的】为早实核桃高效栽培管理提供参考。【方法】以‘中核短枝’和‘新新2号’2个早实核桃品种为研究对象,在种仁油脂转化期到成熟期每隔10 d进行取样,测定种仁中粗脂肪、粗蛋白、单宁、总酚、总糖和各脂肪酸组分含量,分析基础营养物质含量和油脂含量的变化动态及二者之间的相关性。【结果】2个早实核桃品种种仁中油脂积累的关键时期是7月下旬—9月中下旬,成熟时‘中核短枝’种仁脂肪含量为67%,‘新新2号’种仁脂肪含量为65%。在2个早实核桃品种种仁中共检测出5种主要脂肪酸组分,分别是棕榈酸、硬脂酸、油酸、亚油酸、α-亚麻酸,‘中核短枝’和‘新新2号’种仁中不饱和脂肪酸含量占总脂肪酸含量的比例分别高达89.60%和88.97%。不饱和脂肪酸的主要成分为油酸和亚油酸,‘中核短枝’和‘新新2号’种仁中油酸的占比分别为21.36%、14.09%,亚油酸的占比分别为58.38%、61.80%。在种仁油脂积累过程中,基础营养物质对脂肪积累也有重要作用,蛋白质含量与脂肪含量极显著负相关,在油脂转化期蛋白质、总糖的消耗量较大,总酚和单宁与不饱和脂肪酸合成密切相关。【结论】‘中核短枝’和‘新新2号’2个早实核桃种仁的油脂积累和脂肪酸组分含量的变化趋势基本一致,在7月下旬—9月中下旬迅速增加。2个早实核桃品种种仁中不饱和脂肪酸含量均较高,主要成分为油酸和亚油酸。‘中核短枝’成熟种仁中脂肪含量较高、单宁含量和总酚含量较低,可作为优质核桃品种进行栽培利用。

Top-grain filling characteristics at an early stage of maize(Zea mays L.) with different nitrogen use efficiencies

Maize genotypes vary significantly in their nitrogen use efficiencies（NUEs）.Better understanding of early grain filling characteristics of maize is important,especially for maize with different NUEs.The objectives of this research were（i）to investigate the difference in apical kernel development of maize with different NUEs,（ii）to determine the reaction of apical kernel development to N application levels,and（iii）to evaluate the relationship between apical kernel development and grain yield（GY）for different genotypes of maize.Three maize hybrid varieties with different NUEs were cultivated in a field with different levels of N fertilizer arranged during two growing seasons.Kernel fresh weight（KFW）,volume（KV）and dry weight（KDW）of apical kernel were evaluated at an early grain filling stage.Ear characteristics,GY and its components were determined at maturity stage.Apical kernel of the high N and high efficiency（HN-HE）type（under low N,the yield is lower,and under higher N,the yield is higher）developed better under high N（N210 and N240,pure N of 210 and 240 kg ha^–1）than at low N（N120 and N140,pure N of 120 and 140 kg ha^–1）.The low N and high efficiency（LN-HE）type（under low N,the yield is higher,while under higher N,the yield is not significantly higher）developed better under low N than at high N.The double high efficiency（D-HE）type（for both low and high N,the yield is higher）performed well under both high and low N.Apical kernel reacted differently to the N supply.Apical kernel developed well at an early grain filling stage and resulted in a higher kernel number（KN）,kernel weight（KW）and GY with better ear characteristics at maturity.

The U6 Biogenesis-Like I Plays an Important Role in Maize Kernel and Seedling Development by Affecting the 3＇ End Processing of U6 snRNA

Regulation of gene expression at the post-transcriptional level is of crucial importance in the development of an organism. Here we present the characterization of a maize gene, U6 biogenesis-like 1 （UBL1）, which plays an important role in kernel and seedling development by influencing pre-mRNA splicing. The ubll mutant, exhibiting small kernel and weak seedling, was isolated from a Mutator-tagged population. Trans- genic complementation and three independent mutant alleles confirmed that UBL1, which encodes a putative RNA exonuclease belonging to the 2H phosphodiesterase superfamily, is responsible for the phenotype of ubll. We demonstrated that UBL1 possess the RNA exonuclease activity in vitro and found that loss of UBL1 function in ubll causes decreased level and abnormal 3＇ end constitution of snRNA U6, resulting in splicing defect of mRNAs. Through the in vitro and in vivo studies replacing two histidines with alanines in the H-X-T/S-X （X is a hydrophobic residue） motifs we demonstrated that these two motifs are essential for the normal function of UBL1. We further showed that the function of UBL1 may be conserved across a wide phylogenetic distance as the heterologous expression of maize UBL1 could complement the Arabidopsis ubll mutant.

不同发育时期小麦粒重性状QTL的动态分析

利用6044×01-35构建的重组自交系(RIL)群体为试验材料,对小麦粒重性状进行发育动态QTL分析。结果表明,在小麦花后子粒灌浆的7个不同时期,两个试验点共检测到16个与粒重性状相关的QTL。其中开花后20d检测到的单穗粒重QTL位于2A染色体上,解释率达12%,遗传效应超过10;两环境下控制千粒重QTL在7个时期均被检测到。花后的各个时期均能在Xgwm448-Xgpw7399标记区间定位到千粒重QTL。其中花后10d检测到1个千粒重QTL,位于2A染色体的Xgwm448-Xgpw7399标记区间,解释较大的表型变异,达到18%。Qtl8、Qtl13和Qtl14均定位在Xgwm448-Xgpw7399标记区间的同一位置,共同解释11%的表型变异。花后20d和花后25d均检测到1个QTL,位于2A染色体的Xgwm372-Xgwm95标记区间的不同位点,均能解释4%的表型变异。花后40d检测到1个QTL,位于1D染色体的Xwmc93-Xgpw2224标记区间,解释1%的表型变异。从连锁群的位置上看,控制千粒重的QTL主要集中在2A染色体的Xgwm448-Xgpw7399标记区间,这是一个控制千粒重QTL的富集区域,以期进行精细定位和图位克隆。

Identification of genes involved in the formation of soluble dietary fiber in winter rye grain and their expression in cultivars with different viscosities of wholemeal water extract

The grain of rye(Secale cereale L.) used for baking contains a large amount of non-starch polysaccharides,making it an excellent component of functional foods. But rye grain intended for alcohol production and forage use should have a reduced content of these polysaccharides. A comprehensive parameter that can predict the best field of application for winter rye grain is the viscosity of its wholemeal water extract.However, our understanding of the genetic background underlying this key trait and associated features of rye grain is poor. By analyzing six Russian winter rye cultivars, we identified the most contrasting forms and characterized the peculiarities of their water-soluble carbohydrates capable of influencing the viscosity of water extracts. Then, using phylogenetic and transcriptomic analyses, we identified in the rye genome many genes encoding putative glycosyltransferases and glycosylhydrolases responsible for the synthesis and degradation of arabinoxylans, mixed-linkage glucans, cellulose, and some other polysaccharides. We determined the dynamics of m RNA abundance for these genes at three stages of kernel development. Comparisons of gene expression levels in two contrasting cultivars revealed specific members of multigene families that may serve as promising targets for manipulating non-starch polysaccharide content in rye grain. High-viscosity cultivars were characterized by up-regulation of many glycosyltransferases involved in the biosynthesis of arabinoxylans and other cell-wall polysaccharides,whereas low-viscosity cultivars showed up-regulation of several genes encoding polysaccharidedegrading enzymes.

Defective Kernel 39 encodes a PPR protein required for seed development in maize

RNA editing is a posttranscriptional process that is important in mitochondria and plastids of higher plants. All RNA editing-specific trans-factors reported so far belong to PLS-class of pentatricopeptide repeat(PPR)proteins. Here, we report the map-based cloning and molecular characterization of a defective kernel mutant dek39 in maize. Loss of Dek39 function leads to delayed embryogenesis and endosperm development, reduced kernel size, and seedling lethality. Dek39 encodes an E subclass PPR protein that targets to both mitochondria and chloroplasts, and is involved in RNA editing in mitochondrial NADH dehydrogenase3(nad3) at nad3-247 and nad3-275. C-to-U editing of nad3-275 is not conserved and even lost in Arabidopsis, consistent with the idea that no close DEK39 homologs are present in Arabidopsis. However, the amino acids generated by editing nad3-247 and nad3-275 are highly conserved in many other plant species, and the reductions of editing at these two sites decrease the activity of mitochondria NADH dehydrogenase complex I,indicating that the alteration of amino acid sequence is necessary for Nad3 function. Our results indicate that Dek39 encodes an E sub-class PPR protein that is involved in RNA editing of multiple sites and is necessary for seed development of maize.

The Role of PIN Auxin Efflux Carriers in Polar Auxin Transport and Accumulation and Their Effect on Shaping Maize Development

In plants, proper seed development and the continuing post-embryonic organogenesis both require that dif- ferent cell types are correctly differentiated in response to internal and external stimuli. Among internal stimuli, plant hormones and particularly auxin and its polar transport （PAT） have been shown to regulate a multitude of plant phys- iological processes during vegetative and reproductive development. Although our current auxin knowledge is almost based on the results from researches on the eudicot Arabidopsis thaliana, during the last few years, many studies tried to transfer this knowledge from model to crop species, maize in particular. Applications of auxin transport inhibitors, mutant characterization, and molecular and cell biology approaches, facilitated by the sequencing of the maize genome, allowed the identification of genes involved in auxin metabolism, signaling, and particularly in polar auxin transport. PIN auxin efflux carriers have been shown to play an essential role in regulating PAT during both seed and post-embryonic development in maize. In this review, we provide a summary of the recent findings on PIN-mediated polar auxin transport during maize development. Similarities and differences between maize and Arabidopsis are analyzed and discussed, also considering that their different plant architecture depends on the differentiation of structures whose development is con- trolled by auxins.