Cloning, Characterization and Transformation of Methyltransferase 2a Gene (Zmet2a) in Maize (Zea mays L.)

DNA methylation is an important epigenetic regulatory mechanism,it regulates gene expression by recruiting proteins involved in gene repression or by inhibiting the binding of transcription factor(s)to DNA.In this study,a novel methyltransferase 2a gene(Zmet2a)was cloned in maize and identified by polymerase chain reaction-base(PCR-base)using a bioinformatics strategy.The Zmet2a cDNA sequence is 2739 bp long and translates to 912 amino acid peptides.The Zmet2a protein revealed that it contains BAH and CHROMO structural domains,is a non-transmembrane protein that is hydrophilically unstable,and has no signal peptide structure.Meanwhile,we verified the biological roles of Zmet2a using transgenic Arabidopsis overexpressing Zmet2a and Zmet2a-knockout maize.Transgenic Zmet2a Arabidopsis thaliana showed highly significant advancement inflowering time,and Zmet2a-knockout maize showed advancement inflowering time,with significant changes in several traits.Altogether,these report the role of Zmet2a in the regulation offlowering time,which will lay a foundation for revealing the biological function and epigenetic regulation mechanism of Zmet2a in the growth,development andflowering of maize.

Genetic and Agronomic Parameter Estimates of Growth, Yield and Related Traits of Maize (Zea mays L.) under Different Rates of Nitrogen Fertilization

This study evaluated the genetic and agronomic parameter estimates of maize under different nitrogen rates. The trial was established at the Njala Agricultural Research Centre experimental site during 2021 and 2022 in a split block design with three maize varieties (IWCD2, 2009EVDT, and DMR-ESR-Yellow) and seven nitrogen (0, 30, 60, 90, 120, 150 and 180 kg∙N∙ha−1) rates. Findings showed that cob diameter and anthesis silking time (ASI) had intermediate heritability, ASI had high genetic advance, ASI and grain yield had high genotypic coefficient of variation (GCV), while traits with high phenotypic coefficient of variation (PCV) were plant height, ASI, grain yield, number of kernel per cob, number of kernel rows, ear length, and ear height. The PCV values were higher than GCV, indicating the influence of the environment in the studied traits. Nitrogen rates and variety significantly (p < 0.05) influenced grain yield production. Mean grain yields and economic parameter estimates increased with increasing nitrogen rates, with the 30 and 180 kg∙N∙ha−1 plots exhibiting the lowest and highest grain yields of 1238 kg∙ha−1 and 2098 kg∙ha−1, respectively. Variety and nitrogen effects on partial factor productivity (PFPN), agronomic efficiency (AEN), net returns (NR), value cost ratio (VCR) and marginal return (MR) indicated that these parameters were significantly affected (p < 0.05) by these factors. The highest PFPN (41.3 kg grain kg−1∙N) and AEN (29.4 kg grain kg−1∙N) were obtained in the 30 kg∙N∙ha−1 plots, while the highest VCR (2.8) and MR (SLL 1.8 SLL−1 spent on N) were obtained in the 180 kg∙N∙ha−1. The significant influence of variety and nitrogen on traits suggests that increasing yields and maximizing profits require use of appropriate nitrogen fertilization and improved farming practices that could be exploited for increased productivity of maize.

密度与氮肥对关中灌区夏玉米(Zea mays L.)群体光合生理指标的影响

合理的密度是玉米构建良好群体结构、优化群体光合生理功能的基础,适宜的氮肥施用量是玉米进行光合生产的营养物质保障。运用作物群体生理学的方法,采用二次饱和D试验设计,研究了关中灌区夏玉米密度与氮肥耦合和群体光合生理指标的关系及其效应,明确了在关中灌区夏播条件下,在45000-75000株/hm^2的密度范围内,密度与玉米籽粒产量、总干物质积累量（DMA）、吐丝期叶面积指数（MLAI）、总光合势（LAD）、平均作物生长率（CGR）为正相关,在45000-60000株/hm^2的密度范围内,密度与玉米叶片平均净同化率（NAR）为正相关,而在60000-75000株/hm^2的密度范围内,密度与玉米NAR为负相关;在氮素施用量≤600.0 kg/hm^2的范围内,氮素施用量与玉米籽粒产量、DMA、CGR为正相关,在氮素施用量≤260.55 kg/hm^2的范围内,氮素施用量与玉米MLAI、LAD为正相关,与玉米NAR为负相关,而在氮素施用量260.55-600.0kg/hm^2的范围内,氮素施用量与玉米MLAI、LAD为负相关,与玉米NAR为正相关。密度对其影响较大的指标为：籽粒产量、DMA、LAD、CGR和MLAI,影响较小的指标为：NAR;氮肥对其影响较大的指标为：DMA、CGR、籽粒产量、NAR,影响较小的指标为：LAD和MLAI。对籽粒产量和群体生理指标的综合影响效应,密度显著大于氮肥,玉米生产中,尤其要重视合理密植。通过对回归模型的解析,筛选确定了陕单8806玉米在关中灌区夏播中,实现高产的密度与氮肥耦合优化技术方案：合理密度为61713-66177株/hm^2,适宜纯氮施用量为309.88-569.02kg/hm^2。

麦茬处理方式对夏玉米(Zea mays L.)根际生物活性的影响

通过两年3点的大田试验,研究了不同麦茬处理方式(平茬、立茬、除茬)对夏玉米(Zea mays L.)根际生物活性的影响。结果表明:不同麦茬处理的夏玉米根际微生物数量、土壤蛋白酶和脲酶活性均呈先升后降的变化趋势,且在吐丝期达到最大值,但在不同生育时期,不同处理间有所差异,趋势为平茬>立茬>除茬,且苗期差异较大,后期逐渐减小。麦茬处理方式对夏玉米田土壤碳通量的影响也表现出同样趋势。

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.

Screening Methods for Waterlogging Tolerance at Maize (Zea mays L.) Seedling Stage

Waterlogging strongly affects agronomic performance of maize （Zea mays L.）. In order to investigate the suitable selection criteria of waterflooding tolerant genotypes, and identify the most susceptible stage and the best continuous treatment time to waterlogging, 20 common maize inbred lines were subjected to successive artificial waterflooding at seedling stage, and waterlogging tolerance coefficient （WTC） was used to screen waterflooding tolerant genotypes. In addition, peroxidase （POD） activities and malondialdehyde （MDA） contents were measured for 6 of 20 lines. The results showed that the second leaf stage （V2） was the most susceptible stage, and 6 d after waterflooding was the best continuous treatment time. Dry weight （DW） of both shoots and roots of all lines were significantly reduced at 6 d time-point of waterlogging, compared to control. POD activities and MDA contents were negatively and significantly correlated, and the correlation coefficient was -0.9686 （P 〈 0.0001）. According to the results, WTC of shoot DW can be used for practical screening as a suitable index, which is significantly different from control and waterlogged plants happened 6 d earlier. Furthermore, leaf chlorosis, MDA content and POD activities could also be used as reference index for material screening. The implications of the results for waterlogging-tolerant material screening and waterlogging-tolerant breeding have been discussed in maize.

常乐稀土复合肥对玉米(Zea mays L.)的细胞毒性和遗传毒性效应

为了研究常乐稀土多元复合肥对主要农作物--玉米(Zea mays L.)是否产生细胞毒性和遗传毒性效应,采用不同梯度质量浓度的稀土复合肥溶液对玉米进行处理,观察其根尖生长状况,统计分析细胞死亡率、微核率、有丝分裂指数以及对细胞DNA的损伤情况.结果表明,质量浓度为10mg·L-1以下时,常乐稀土复合肥能够促进玉米根尖的生长,100mg·L-1以上时抑制根尖生长;随着试验浓度的递增,细胞死亡率、微核率、DNA损伤率逐渐上升并具有正相关性,表现出明显的剂量-效应关系,而有丝分裂指数却随着浓度的增加逐渐下降.以上结果表明,常乐稀土复合肥对玉米确有一定的细胞毒性和遗传毒性效应,其细胞毒性阈值和遗传毒性阈值皆为100mg·L-1.

Activity of Acetolactate Synthase from Maize (Zea mays L.) as Influenced by Chlorsulfuron and Tribenuron-methyl

Study on relative sensitivity of maize (Zea mays L.) Nongda108 and Nongda3138 to sulfony-lurea herbicide chlorsulfuron and tribenuron-methyl using maize taproot length by sand bioassy indicated that, Nongda3138 had higher tolerance to chlorsulfuron and tribenuron-methyl than Nongda108 did. Chlorsulfuron had stronger growth inhibition to maize Nongda108 and Nongda3138 than tribenuron-methyl did. Study on target enzyme of sulfonylurea herbicide acetolactate synthase (ALS) showed that, chlorsulfuron and tribenuron-methyl inhibited ALS in vitro strongly, and non-competitively. In the same concentration of inhibitors, chlorsulfuron had stronger ALS activity inhibition than tribenuron-methyl did. Lower level of chlorsulfuron and tribenuron-methyl has no ALS activity inhibition in vivo, the ALS inhibition only occurred in the condition of high concentration of chlorsulfuron and tribenuron-methyl in vivo.

Genetic dissection of husk number and length across multiple environments and fine-mapping of a major-effect QTL for husk number in maize(Zea mays L.)

Husk number(HN)and husk length(HL)influence the mechanical harvesting of maize grain.We investigated the genetic basis of HN and HL using a population of 204 recombinant inbred lines phenotypically evaluated in five environments.The two husk traits showed broad phenotypic variation and high heritability.Nine stable quantitative trait loci(QTL)were identified by single-environment mapping,comprising four QTL for HN and five for HL,and three QTL explained>10%of the phenotypic variation.Joint mapping revealed 22 additive QTL and 46 epistatic QTL.Both additive and epistatic(additive×additive)effects as well as a few large-effect QTL and some minor-effect QTL appeared to contribute to the genetic architecture of HN and HL.The QTL for HN located on chromosome 7,q HN7,which accounted for^20%of phenotypic variation,was detected in all five environments.q HN7 was fine-mapped to a 721.1 kb physical region based on the maize B73 Ref Gen_v3 genome assembly.Within this interval,four genes associated with plant growth and development were selected as candidate genes.The results will be useful for improvement of maize husk traits by molecular breeding and provide a basis for the cloning of q HN7.

QTL analysis of the developmental changes in cell wall components and forage digestibility in maize(Zea mays L.)

Cell wall architecture plays a key role in stalk strength and forage digestibility.Lignin,cellulose,and hemicellulose are the three main components of plant cell walls,and they can impact stalk quality by affecting the structure and strength of the cell wall.To explore cell wall development during secondary cell wall lignification in maize stalks,conventional and conditional genetic mapping were used to identify the dynamic quantitative trait loci(QTLs)of the cell wall components and digestibility traits during five growth stages after silking.Acid detergent lignin(ADL),cellulose(CEL),acid detergent fiber(ADF),neutral detergent fiber(NDF),and in vitro dry matter digestibility(IVDMD)were evaluated in a maize recombinant inbred line(RIL)population.ADL,CEL,ADF,and NDF gradually increased from 10 to 40 days after silking(DAS),and then they decreased.IVDMD initially decreased until 40 DAS,and then it increased slightly.Seventytwo QTLs were identified for the five traits,and each accounted for 3.48–24.04%of the phenotypic variation.Six QTL hotspots were found,and they were localized in the 1.08,2.04,2.07,7.03,8.05,and 9.03 bins of the maize genome.Within the interval of the pleiotropic QTL identified in bin 1.08 of the maize genome,six genes associated with cell wall component biosynthesis were identified as potential candidate genes for stalk strength as well as cell wall-related traits.In addition,26 conditional QTLs were detected in the five stages for all of the investigated traits.Twenty-two of the 26 conditional QTLs were found at 30 DAS conditioned using the values of 20 DAS,and at 50 DAS conditioned using the values of 40 DAS.These results indicated that cell wall-related traits are regulated by many genes,which are specifically expressed at different stages after silking.Simultaneous improvements in both forage digestibility and lodging resistance could be achieved by pyramiding multiple beneficial QTL alleles identified in this study.

Establishment and Optimization of the Regeneration System of Mature Embryos of Maize (Zea mays L.)

A reliable system was developed for regeneration from mature embryos derived from callus of four maize inbred lines （Liao 7980, Dan 9818, Dan 340, and Dan 5026）. The protocol was mainly based on a series of experiments involving the composition of culture medium. We found that 9 pM 2,4-dichlorophenoxyacetic acid in MS medium was optimum for the induction of callus. The induction frequency of primary calli was over 85% for four inbred lines tested. The addition of L- proline （12 mM） in subculture medium significantly promoted the formation of embryogenic callus but it did not significantly enhance growth rate of callus. Efficient shoot regeneration was obtained on regeneration medium containing 2.22 μM 6- benzylaminopurine in combinations with 4.64 μM Kinetin. Regenerated shoots were rooted on half-strength MS medium containing 2.85 μM indole-3-butyric acid. This plant regeneration system provides a foundation for genetic transformation of maize.

玉米(Zea mays L.)×大刍草(Zea diploperennis L.)远缘杂交选育玉米自交系的研究

为了丰富玉米遗传变异,从1990年起,我们进行了玉米(Zea mays L.)和多年生二倍体大刍草(Zea diploperennis L.)远缘杂交的研究。经过5年8个世代的杂交、回交和自交选择,目前已获得抗逆性、抗病虫能力较强,农艺性状优良的玉米自交系14个。在测配1000个杂交组合中,所选系表现出较高的一般配合力和特殊配合力。实践证明,这是一条导入外缘种质、拓宽玉米遗传基础,进一步提高杂交种增产潜力的途径之一。

Genomic and transcriptomic insights into cytochrome P450 monooxygenase genes involved in nicosulfuron tolerance in maize(Zea mays L.)

Postemergence application of nicosulfuron for weed control in maize fields can cause great damage to certain maize inbred lines and hybrids. Two maize genotypes, tolerant inbred(HBR) and sensitive inbred(HBS), were found to significantly differ in their phenotypic responses to nicosulfuron, with the EC50(50% effective concentration) values differed statistically(763.6 and 5.9 g a.i. ha–1, respectively). Pre-treatment with malathion, a known cytochrome P450 inhibitor, increased nicosulfuron injury in both HBR and HBS. Our results support the hypothesis that nicosulfuron selectivity in maize is associated with cytochrome P450 metabolism. Further analysis of the maize genome resulted in the identification of 314 full length cytochrome P450 monooxygenase(CYP) genes. These genes were classified into 2 types, 10 clans and 44 families. The CYP71 clan was represented by all A-type genes(168) belonging to 17 families. Nine clans possessed 27 families containing 146 non-A-type genes. The consensus sequences of the heme-binding regions of A-type and non-A-type CYP proteins are ‘PFGXGRRXCPG’ and ‘FXXGPRXCXG’, respectively. Illumina transcriptome sequence results showed that there were 53 differentially expressed CYP genes on the basis of high variation in expression between HBS and HBR, nicosulfuron-treated and untreated samples. These genes may contribute to nicosulfuron tolerance in maize. A hierarchical clustering analysis obtained four main clusters named C1 to C4 in which 4, 15, 21, and 13 CYP genes were found in each respective cluster. The expression patterns of some CYP genes were confirmed by RT-q PCR analysis. The research will improve our understanding of the function of maize cytochrome P450 in herbicide metabolism.

Ameliorative Effects of Brassinosteroid on Excess Manganese-Induced Oxidative Stress in Zea mays L. Leaves

Manganese （Mn） is becoming an important factor limiting crop growth and yields especially on acid soils. The present study was designed to explore the hypothesis that brassinosteroid application can enhance the tolerance of maize （Zea mays L.） to Mn stress and if so, whether or not the mechanism underlying involves regulation of antioxidative metabolism in leaves. The effects of 24-epibrassinosteroid （EBR） on the growth, photosynthesis, water status, lipid peroxidation, accumulation of reactive oxygen species, and activities or contents of antioxidant defense system in maize plants under Mn stress were investigated by a pot experiment. At supplemented Mn concentrations of 150-750 mg kg^-1 soil, the growth of plants was inhibited in a concentration-dependent manner. The semi-lethal concentration was 550 mg Mn kgq soil. Foliage application with 0.1 mg L^-1 EBR significantly reduced the decrease in dry mass, chlorophyll content, photosynthetic rate, leaf water content, and water potential of plants grown in the soil spiked with 550 mg kg^-1 Mn. The oxidative stress caused by excess Mn, as reflected by the increase in malondialdehyde （MDA） content and lipoxygenase （LOX, EC 1.13.11.12） activity, accumulation of superoxide radical and H2O2, was greatly decreased by EBR treatment. Further investigations revealed that EBR application enhanced the activities of superoxide dismutase （SOD, EC 1.15.1.1）, peroxidase （POD, EC 1.11.1.7）, catalase （EC 1.11.1.6）, ascorbate peroxidase （APX, EC 1.11. 1.11）, dehydroascorbate reductase （DHAR, EC 1.8.5.1）, and glutathione reductase （GR, EC 1.6.4.2）, and the contents of reduced ascorbate and glutathione, compared with the plants without EBR treatment. It is concluded that the ameliorative effects of EBR on Mn toxicity are due to the upregulation of antioxidative capacity in maize under Mn stress.

Breeding for Drought Tolerance in Maize (Zea mays L.)

Drought, like many other environmental stresses, has adverse effects on crop yield including maize (Zea mays L.). Low water availability is one of the major causes for maize yield reductions affecting the majority of the farmed regions around the world. Therefore, the development of drought-tolerant lines becomes increasingly more important. In maize, a major effect of water stress is a delay in silking, resulting in an increase in the anthesis-silking interval, which is an important cause of yield failures. Diverse strategies are used by breeding programs to improve drought tolerance. Conventional breeding has improved the drought tolerance of temperate maize hybrids and the use of managed drought environments, accurate phenotyping, and the identification and deployment of secondary traits has been effective in improving the drought tolerance of tropical maize populations and hybrids as well. The contribution of molecular biology will be potential to identify key genes involved in metabolic pathways related to the stress response. Functional genomics, reverse and forward genetics, and comparative genomics are all being deployed with a view to achieving these goals. However, a multidisciplinary approach, which ties together breeding, physiology and molecular genetics, can bring a synergistic understanding to the response of maize to water deficit and improve the breeding efficiency.

Modification of total and phosphorus mineralizing bacterial communities associated with Zea mays L.through plant development and fertilization regimes

Harnessing the rhizospheric microbiome,including phosphorus mineralizing bacteria(PMB),is a promising technique for maintaining sustainability and productivity in intensive agricultural systems.However,it is unclear as to which beneficial taxonomic group populations in the rhizosphere are potentially associated with the changes in soil microbiomes shifted by fertilization regimes.Herein,we analyzed the diversity and community structure of total bacteria and PMB in the rhizosphere of maize(Zea mays L.)grown in soils under 25 years of four fertilization regimes(compost,biocompost,chemical,or nonfertilized)via selective culture and Illumina sequencing of the 16S rRNA genes.Plant development explained more variations(29 and 13%,respectively)in the composition of total bacteria and PMB in the rhizosphere of maize than the different fertilization regimes.Among those genera enriched in the rhizosphere of maize,the relative abundances of Oceanobacillus,Bacillus,Achromobacter,Ensifer,Paracoccus,Ramlibacter,and Luteimonas were positively correlated with those in the bulk soil.The relative abundance of Paracoccus was significantly higher in soils fertilized by compost or biocompost than the other soils.Similar results were also observed for PMB affiliated with Ensifer,Bacillus,and Streptomyces.Although plant development was the major factor in shaping the rhizospheric microbiome of maize,fertilization regimes might have modified beneficial rhizospheric microbial taxa such as Bacillus and Ensifer.

A new flavonoid from the bract of Zea mays L.

A new flavonoid was isolated from the bract of Zea mays L.The structure of the compound was identified as 4＇,5,7-trihydroxy-3＇,5＇-dimethoxyflavone 7-O-[β-D-apiofuranosyl（1→2）]-β-D-glucopyranoside on the ground of chemical and spectroscopic methods.

Study on the Photosynthetic Characteristics in Spring Maize (Zea mays L.)of Different Quality Type

common maize synthetic rate（ Photosynthetic characteristics were probed by sweet maize, waxy maize, high starch maize and The results revealed that leaf area index （LAI）, chlorophyll a content, chlorophyll b content,photo-PR） showed single peak curve at the whole growth stage. The stages of peak were different according to different varieties. NEAUS4 had the lowest peak and while SIDAN 19 had the highest among all stages. Ratio of chlorophyll a to b was low at seedling stage, reached the peak atjointing stage and then declined. SIDAN 19 had the lower level at the last stages.

Influence of Soil Moisture and Air Temperature on the Stability of Cytoplasmic Male Sterility (CMS) in Maize (Zea mays L.)

Cytoplasmic male sterility (CMS) is a maternally inherited trait that suppresses the production of viable pollen. CMS is a useful biological tool for confinement strategies to facilitate coexistence of genetically modified (GM) and non-GM crops in case where it is required. The trait is reversible and can be restored to fertility in the presence of nuclear restorer genes (Rf genes) and by environmental impacts. The aim of this study was to investigate the influence of the level of irrigation on the stability of CMS maize hybrids under defined greenhouse conditions. Additionally the combination of irrigation and air temperature was studied. Three CMS maize hybrids were grown with different levels of irrigation and in different temperature regimes. Tassel characteristics, pollen production and fertility were assessed. The CMS stability was high in hot air temperatures and decreased in lower temperatures. The level of irrigation had no major effect on the level of sterility. The extent of these phenomena was depending on the genotype of CMS maize and should be known before using CMS for coexistence purposes.

玉米(Zea mays L.)叶脉发育的研究

玉米的叶脉在单子叶植物中有一定的代表性，叶脉由四级组成，粗细不同的一、二、三级叶脉均从叶基向叶尖方向延伸，属叶片的纵向叶脉，四级脉横向与一、二、三级叶脉连接，是横向的叶脉，各级叶脉有各自的形成方式，由于它们有规律的分布，从而构成了叶片的输导网络，各级叶脉的发生和发育与叫片的生长有直接的关系。