Variations in chlorophyll content, stomatal conductance, and photosynthesis in Setaria EMS mutants

Chlorophyll (Chl) content,especially Chl b content,and stomatal conductance (G_s) are the key factors affecting the net photosynthetic rate (P_n).Setaria italica,a diploid C_4 panicoid species with a simple genome and high transformation efficiency,has been widely accepted as a model in photosynthesis and drought-tolerance research.The current study characterized Chl content,G_s,and P_n of 48 Setaria mutants induced by ethyl methanesulfonate.A total of 24,34,and 35 mutants had significant variations in Chl content,G_s,and P_n,respectively.Correlation analysis showed a positive correlation between increased G_s and increased P_n,and a weak correlation between decreased Chl b content and decreased P_n was also found.Remarkably,two mutants behaved with significantly decreased Chl b content but increased P_n compared to Yugu 1.Seven mutants behaved with significantly decreased G_s but did not decrease P_(n )compared to Yugu 1.The current study thus identified various genetic lines,further exploration of which would be beneficial to elucidate the relationship between Chl content,G_s,and P_n and the mechanism underlying why C_4 species are efficient at photosynthesis and water saving.

Generation of oxidants and their role in the metabolism of oxygen in the survivability of filarial parasite Setaria digitata in cattle:a possible intervention strategy

Background:The goal of the current study was to identify the potential therapeutic targets by analyzing the production of oxidants and their effects on the antioxidant system of Setaria digitata,a filarial worm that was collected from the peritoneal cavity of recently butchered calves.Methods:S.digitata was collected and the mitochondria-like particles(MLP),somatic isolates and the post-nuclear fraction were prepared for investigation.The radical intensity and the antioxidant enzyme activity in these fractions were evaluated.The effect of diethylcarbamazine(DEC),Q0,omeprazole,and L-methyl arginine on NOS production in the worm was also evaluated.Results:Experiments showed that coelomic fluid and whole worm showed significant levels of Nitric oxide compared to other fractions.The cytosol of the worm showed higher NOS activity than MLP.Superoxide dismutase and nitric oxide(NO)levels were found to be higher in MLP fraction of the worm.It was also found that DEC did not have any effect on the prevention of NO,while Q0,omeprazole,and L-methyl arginine treatment showed a decrease in NO production significantly.Conclusion:The results showed that by using the existing drugs we could reduce the propagation of S.digitata by inhibiting the production of NO.

谷子(Setaria italica)Ch 4n×法氏狗尾草(S.faberii)杂种幼胚培养及杂种F_1的鉴定

谷子是我国北方主要作物之一,耐干旱和瘠薄,在西北和华北干旱地区广为栽培,但产量较低。我们在谷子三系选育研究中,利用远缘杂交,以狗尾草属的5个野生种(包括法氏狗尾草在内)为母本与谷子杂交,分别选育不育系,1989年安排其反交组合 Ch 4n×法氏狗尾草,其目的在于:(一)根据雄性不育基因的多样性,彼此独立性和质核对应性原理,通过正反交对比,测验此雄性不育基因有无互补关系;(二)

增强UV-B辐射与不同水平氮素对谷子(Setaria italica(L．)Beauv．)叶片保护物质及保护酶的影响

研究了生长在1.875 mmol.L-1和15 mmol.L-1硝态氮素水平条件下的谷子(Setaria italica(L.)Beauv.)开花期间在进行强度为7.12 kJ.m-.2d-1增强UV-B辐射处理时叶片类黄酮含量、苯丙氨酸解氨酶(PAL)和保护酶活性变化差异。主要结果表明在开花期无论是否进行增强UV-B辐射处理,较低水平氮素均比较高水平氮素更有利于提高谷子叶片PAL活性;叶片类黄酮含量除在进行增强UV-B处理时较低氮素条件下生长的谷子在开花末期显著高于较高氮素条件下生长的谷子外,受氮素水平影响不甚明显。而在开花期不进行与进行增强UV-B辐射处理,氮素水平对叶片保护酶的影响有所差异:不进行增强UV-B辐射处理,整个开花期氮素水平对谷子叶片SOD活性有显著影响而对ASP活性无显著影响,对CAT和POD活性则在开花期部分阶段有显著影响。进行增强UV-B辐射处理,整个开花期氮素水平对谷子叶片SOD与CAT活性有显著影响而对ASP、POD活性影响不显著。

古粟(Setaria italica Beauv.)研究综述

粟于中国北方俗称"谷子",脱壳后称"小米",南方则通称"小米",在古代粮食作物中的地位举足轻重。近世学者对粟的史前考古发现、起源与传播、稷的粟黍之争、生产与文化等方面行了研究,并且已经取得了很多成果,但在某些方面的探索还非常不够,如史前人类生活中粟作的地位与演变;古粟的栽培、储藏、加工与利用技术;历史上粟的生产布局、价格与产量的变动;粟与古代经济、政治、文化的关系等。今后应加强在这些方面的投入,不断拓展粟研究的深度和广度。

Genetic Analysis and Preliminary Mapping of a Highly Male-Sterile Gene in Foxtail Millet(Setaria italica L.Beauv.) Using SSR Markers

Breeding of male-sterile lines has become the mainstream for the heterosis utilization in foxtail millet,but the genetic basis of most male-sterile lines used for the hybrid is still an area to be elucidated.In this study,a highly male-sterile line Gao146A was investigated.Genetic analysis indicated that the highly male-sterile phenotype was controlled by a single recessive gene a single recessive gene.Using F 2 population derived from cross Gao146A/K103,one gene controlling the highly male- sterility,tentatively named as ms1,which linked to SSR marker b234 with genetic distance of 16.7 cM,was mapped on the chromosome VI.These results not only laid the foundation for fine mapping of this highly male-sterile gene,but also helped to accelerate the improvement of highly male-sterile lines by using molecular marker assisted breeding method.

The NAC-like transcription factor Si NAC110 in foxtail millet(Setaria italica L.) confers tolerance to drought and high salt stress through an ABA independent signaling pathway

Foxtail millet（Setaria italica（L.）P.Beauv）is a naturally stress tolerant crop.Compared to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance traits.To date,the scope of functional genomics research in foxtail millet（S.italic L.）has been quite limited.NAC（NAM,ATAF1/2 and CUC2）-like transcription factors are known to be involved in various biological processes,including abiotic stress responses.In our previous foxtail millet（S.italic L.）RNA seq analysis,we found that the expression of a NAC-like transcription factor,SiNAC110,could be induced by drought stress;additionally,other references have reported that SiNAC110 expression could be induced by abiotic stress.So,we here selected SiNAC110 for further characterization and functional analysis.First,the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM（no apical meristem）domain between the 11–139 amino acid positions.Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family.Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts.Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration,high salinity and other abiotic stresses.Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that,under drought and high salt stress conditions,the seed germination rate,root length,root surface area,fresh weight,and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type（WT）,suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses.However,overexpression of SiN AC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid（ABA）treatment.Expression analysis of genes involved in proline synthesis,Na＋/K＋transport,drought responses,and aqueous transport proteins were higher in the SiNAC110overexpressed lines than in the WT,whereas expression of ABA-dependent pathway genes did not change.These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses,likely through influencing the regulation of proline biosynthesis,ion homeostasis and osmotic balance.Therefore,SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.

Heterologous expression,chaperone mediated solubilization and purification of parasitic nematode-specific growth factor-like protein of Setaria digitata

Objective:To clone,express and purify a putative parasitic nematode specific protein of Setaria digitata(S.digitata),filarial nematode that infects livestock and cause significant economic losses in Far East and Asia to he used for structural and functional analyses.Methods:To characterize uneharacterized gene of,S.digitata(SDUG),the herterologous expression of SDUG was carried out in the pET[cloned into pET45b(+)]expression system initially and co-expression of SDUC using chaperoiie plasmids pG-KJE8,pGro 7,pKJE7,pG-Tf2 and pTf16 containing chapcrone proteins of dnaK-dnaJ-grpE-groES-gro-E,groES-groEL,dnaK-dnaJ-grpE,groES-groEL-tig,and tig respectively,was carried out subsequently.Results:Expression of SDUG was seen when Escherichia coli strain BI.21(DE3)is used,while concentrating protein largely into the insoluble fraction.The co-expression of SDUG using chaperoiie plasmid mediated system indicated a significant increase of the protein in the soluble fraction.Of the chaperon plasniid sets,the highest amount of recombinant SDUP in the soluble fraction was seen when pGro7 was used in the presence of2 mg/mL L-arabinosc and 0.6M IPTG concentration in the culture medium and for 3 h of incubation at the temperature of 28℃.Recombinant SDUG was purified both from soluble and insoluble fractions using Ni affinity chromatography.SDS-PAGE and western blot analyses of these proteins revealed a single band having expected size of^24 kDa.Conclusions:SDUG seems to be more aggregate-prone and hydrophobic in nature and such protein can make soluble by correct selecting the inducer concentrations and induction temperature and its duration.

The application of Fourier transform mid-infrared(FTIR) spectroscopy to identify variation in cell wall composition of Setaria italica ecotypes

Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum,maize,sugarcane,switchgrass and Miscanthus×giganteus.Accurate partial least squares regression（PLSR）models to predict S.italica stem composition have been generated,based upon Fourier transform mid-infrared（FTIR）spectra and calibrated with wet chemistry determinations of ground S.italica stem material measured using a modified version of the US National Renewable Energy Laboratory（NREL）acid hydrolysis protocol.The models facilitated a high-throughput screening analysis for glucan,xylan,Klason lignin and acid soluble lignin（ASL）in a collection of 183 natural S.italica variants and clustered them into classes,some possessing unique chemotypes.The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes.Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles,measured by RT-q PCR,among accessions of S.italica,which closely mirror profiles observed in the different developmental regions of an elongating internode of S.viridis by RNASeq.

Effects of Fertility and Density on Biomass Production,Translocation and Lodging Resistance of Millet(Setaria italica L.)in North China

In this study, the plant biomass production, biomass translocation rates across tissues and the lodging resistant-associated traits of millet （ Setaria italica L.） in North China were investigated. Among the four summer millet cultivars, Baogu 19 exhibited improved plant biomass （PB） production at flowering and maturity stages, biomass translocation amount （BTA） from vegetative tissues to seeds during filling period, and lodging resistant-associated （LRA） traits compared with other cultivars, including enhanced stem lignin contents, increased anti-broken resistance （ABR）, anti-puncturing resistance （APR）, and stem diameter （SD） of plants. Compared with treatment regular cultivation （RC）, high fertility treatment （HF） increased the plant BP, BTA from vegetative tissue to seed at filling stage, and the plant LRA traits; whereas high density treatment （HD） decreased the plant BP at plant level, plant BTA from vegetative tissues to seeds at filling stage, and the plant LRA traits. Correlation analysis revealed that stem ABR is significantly correlated with the plant lodging resistant-associated traits including APR and SD in the summer millet cultivars examined under various cultivation treatments. Our investigation indicates that cultivar Baogu 19 together with suitable fertilization and density can promote the plant biomass production, enhance vegetative tissue biomass translocation to seeds, and improve the lodging resistance of summer millet plants in North China.

Calcineurin B-like protein 5(SiCBL5)in Setaria italica enhances salt tolerance by regulating Na^(+)homeostasis

Salinity,a major abiotic stress,reduces plant growth and severely limits agricultural productivity.Plants regulate salt uptake via calcineurin B-like proteins(CBLs).Although extensive studies of the functions of CBLs in response to salt stress have been conducted in Arabidopsis,their functions in Setaria italica are still poorly understood.The foxtail millet genome encodes seven CBLs,of which only SiCBL4 was shown to be involved in salt response.Overexpression of SiCBL5 in Arabidopsis thaliana sos3-1 mutant rescued its salt hypersensitivity phenotype,but that of other SiCBLs(SiCBL1,SiCBL2,SiCBL3,SiCBL6,and SiCBL7)did not rescue the salt hypersensitivity of the Atsos3-1 mutant.SiCBL5 harbors an N-myristoylation motif and is located in the plasma membrane.Overexpression of SiCBL5 in foxtail millet increased its salt tolerance,but its knockdown increased salt hypersensitivity.Yeast two-hybrid and firefly luciferase complementation imaging assays showed that SiCBL5 physically interacted with SiCIPK24 in vitro and in vivo.Cooverexpression of SiCBL5,SiCIPK24,and SiSOS1 in yeast conferred a high-salt-tolerance phenotype.Compared to wild-type plants under salt stress conditions,SiCBL5 overexpressors showed lower accumulations of Na^(+) and stronger Na^(+) efflux,whereas RNAi-SiCBL5 plants showed higher accumulations of Na^(+) and weaker Na^(+) efflux.These results indicate that SiCBL5 confers salt tolerance in foxtail millet by modulating Na^(+) homeostasis.

Identification of no pollen 1 provides a candidate gene for heterosis utilization in foxtail millet(Setaria italica L.)

Male sterility is a common biological phenomenon in plant kingdom and has been used to generate male-sterile lines, which are important genetic resources for commercial hybrid seed production. Although increasing numbers of male-sterility genes have been identified in rice(Oryza sativa) and Arabidopsis(Arabidopsis thaliana), few male-sterility-related genes have been characterized in foxtail millet(Setaria italica). In this study, we isolated a male-sterile ethyl methanesulfonate-generated mutant in foxtail millet, no pollen 1(sinp1), which displayed abnormal Ubisch bodies, defective pollen exine and complete male sterility. Using bulk segregation analysis, we cloned SiNP1 and confirmed its function with CRISPR/Cas9 genome editing. SiNP1 encoded a putative glucose-methanol-choline oxidoreductase.Subcellular localization showed that the SiNP1 protein was preferentially localized to the endoplasmic reticulum and was predominantly expressed in panicle. Transcriptome analysis revealed that many genes were differentially expressed in the sinp1 mutant, some of which encoded proteins putatively involved in carbohydrate metabolism, fatty acid biosynthesis, and lipid transport and metabolism, which were closely associated with pollen wall development. Metabolome analysis revealed the disturbance of flavonoids metabolism and fatty acid biosynthesis in the mutant. In conclusion, identification of SiNP1 provides a candidate male-sterility gene for heterosis utilization in foxtail millet and gives further insight into the mechanism of pollen reproduction in plants.

应用牛腹腔唇乳突丝虫（Setaria labiatopapillosa）抗原的ELISA检测班氏丝虫病人

应用牛腹腔唇乳突丝虫（Ｓｅｔａｒｉａｌａｂｉａｔｏｐａｐｉｌｌｏｓａ）提纯抗原（简称牛丝虫Ｇ抗原）和抗人ＩｇＧ酶标记物的酶联免疫吸附试验，检测２２７例班氏丝虫病人血清的阳性率为９５．５９％；检测１３２例健康人血清的假阳性率为２．２７％，１０例血吸虫病人血清的阳性率为０。应用牛丝虫Ｇ抗原和马来丝虫抗原同步检测５０例班氏丝虫病人血清的阳性率，前者为９８％，后者为８６％，二者有显著差异（ｐ＜０．０５）。因此认为，牛丝虫Ｇ抗原可以作为人丝虫病的异种抗原诊断试剂。

Genetic Diversity and Classification of Chinese Elite Foxtail Millet [Setaria italica (L.) P. Beauv.] Revealed by Acid-PAGE Prolamin

Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.

Effects of fi laricidal drugs on longevity and enzyme activities of the microfilariae of Setaria cervi in white rats

Objective: To analyse the efficacy of diethylcarbamazine(DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microi lariae recovered from the peripheral circulation of the rats before and after the treatment.Methods: Setaria cer vi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microi lariae and dif erential leucocyte counts were recorded till the disappearance of microi lariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microi lariae recovered from normal and treated rats.Results: The microi lariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4-7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most ef ective drug which cleared the microi laraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microi lariae and had no ef ect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.

凯氏丝状线虫(Setaria Kabargi Kadenazzi)在我国的首次发现

本文首次报道寄生于我国野生动物林麝(Moschus berezovskii)腹腔中的凯丝状线虫(Setaria kabargj).

粟蠕孢菌(Helminthosporium setariae=Cochliobolus setariae)氯酸钾抗性突变株的筛选

利用菌株对氯酸钾的抗性,在粟蠕孢菌(Helminthosporium setariae)中筛选氮代谢营养缺陷突变株,获得25株性状稳定的突变菌株。

The m^(6)A reader SiYTH1 enhances drought tolerance by affecting the messenger RNA stability of genes related to stomatal closure and reactive oxygen species scavenging in Setaria italica

Foxtail millet(Setaria italica),a vital drought-resistant crop,plays a significant role in ensuring food and nutritional security.However,its drought resistance mechanism is not fully understood.N6-methyladenosine(m^(6)A)modification of RNA,a prevalent epi-transcriptomic modification in eukaryotes,provides a binding site for m^(6)A readers and affects plant growth and stress responses by regulating RNA metabolism.In this study,we unveiled that the YT521-B homology(YTH)family gene SiYTH1 positively regulated the drought tolerance of foxtail millet.Notably,the siyth1 mutant exhibited reduced stomatal closure and augmented accumulation of excessive H_(2)O_(2)under drought stress.Further investigations demonstrated that SiYTH1 positively regulated the transcripts harboring m^(6)A modification related to stomatal closure and reactive oxygen species(ROS)scavenging under drought stress.SiYTH1 was uniformly distributed in the cytoplasm of SiYTH1-GFP transgenic foxtail millet.It formed dynamic liquid-like SiYTH1 cytosol condensates in response to drought stress.Moreover,the cytoplasmic protein SiYTH1 was identified as a distinct m^(6)A reader,facilitating the stabilization of its directly bound SiARDP and ROS scavenging-related transcripts under drought stress.Furthermore,natural variation analysis revealed SiYTH1AGTG as the dominant allele responsible for drought tolerance in foxtail millet.Collectively,this study provides novel insights into the intricate mechanism of m^(6)A reader-mediated drought tolerance and presents a valuable genetic resource for improving drought tolerance in foxtail millet breeding.

青狗尾草(Setaria viridis)SBP基因家族的鉴定及表达分析

SQUAMOSA启动子结合蛋白家族作为一种转录调控因子广泛存在于植物中,起着调节生长发育及多种生理生化反应。目前,其编码基因已从多种植物中分离鉴定,但是在作为单子叶C4模式生物的青狗尾草中却尚未研究。本研究基于生物信息学方法,从青狗尾草全基因组中鉴定出17个SBP基因,对这些基因的结构、理化性质、染色体定位以及共线性进行分析,并且探讨了这些基因在不同组织的表达情况,此外,还对SBP蛋白质的基序和系统进化进行了分析。结果表明17个SBP基因分布于9条染色体上,其中SvSBP1与SvSBP9、SvSBP3和SvSBP12、SvSBP4和SvSBP14具有线性同源性,共有11个基因包含miR156结合位点。这些基因编码的SBP蛋白表现出不同物种的某些SBP蛋白之间较同一物种的SBP蛋白的遗传关系更近,暗示这些SBP家族成员的基本特征早在单双子叶植物分化之前就已经形成。SvSBP2、SvSBP8、SvSBP13、SvSBP17在所有组织中都有表达,SvSBP15、SvSBP16在所有组织中都不表达外,其余基因均在不同组织中特异性表达,其中花序分生组织中表达的基因数最多,暗示这些基因可能在青狗尾草花的形成中扮演重要角色,并与胁迫响应和叶片的生长发育相关。本研究为进一步鉴定基因功能、阐明青狗尾草生长发育的分子机理提供基础性信息。

谷子萌发期耐盐种质鉴定及应用

本研究利用不同浓度NaCl溶液对10份谷子(Setaria italica L.)种质进行处理,通过分析其萌发期的相对发芽势、相对发芽率、相对芽长以及相对根长等4项指标,明确了适于谷子萌发期耐盐性鉴定的NaCl浓度为180 mmol/L。在该浓度下,利用主成分分析和聚类分析等方法,对180份种质资源进行了耐盐性综合评价和等级划分。结果显示,除相对发芽率和相对芽长之间相关性不显著以外,其余指标之间均呈极显著正相关;主成分分析结果表明,这4项指标可作为谷子耐盐性评价的重要指标;聚类分析结果将180份谷子种质分为极端耐盐、耐盐、盐敏感和极端盐敏感4类;进一步采用隶属函数进行综合评价,筛选到硷谷、衡谷12、齐头白、K-3606和晋谷20等5份极端耐盐种质材料。最后,在该浓度处理下,对黑枝谷×长农35号(极端盐敏感×耐盐)F7代重组近交系遗传群体进行了初步分析,发现40份株系耐盐性等级频率分布近似正态分布,表明该群体适宜耐盐QTL挖掘。研究结果说明,在180 mmol/L NaCl处理下,通过谷子萌发期相对发芽势、相对发芽率、相对芽长和相对根长等4个指标能较好地区分不同种质耐盐性的差异。