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1.
Photosynthesis occurs mainly in chloroplasts, whose development is regulated by proteins encoded by nuclear genes.Among them, pentapeptide repeat(PPR) proteins participate in organelle RNA editing. Although there are more than 450members of the PPR protein family in rice, only a few affect RNA editing in rice chloroplasts. Gene editing technology has created new rice germplasm and mutants, which could be used for rice breeding and gene function study. This study evaluated the functions of OsPPR9...  相似文献   

2.
【目的】叶绿素是参与光合途径最为重要的光合色素。叶绿体的发育及叶绿素的合成在很大程度上依赖于质体基因组与核基因组之间的双向信号传导来精确协调基因表达。通过对白化表型的CRISPR/Cas9-ZmpTAC2转基因阳性纯合突变材料进行RNA-seq研究,筛选和鉴定参与叶绿素合成的相关基因,为明确叶绿素的合成途径奠定基础。【方法】以CRISPR/Cas9-ZmpTAC2玉米转基因编辑纯合突变株系为研究材料,使用透射电镜观察叶绿体超微结构和分光光度法测定叶片叶绿素含量,确定叶绿体发育状态及叶绿素合成情况。对转基因阴性材料（CK）和CRISPR/Cas9-ZmpTAC2转基因纯合编辑材料（zmptac2）苗期叶片取样进行转录组测序。通过生物信息学分析,寻找CK与zmptac2间差异表达的基因;qRT-PCR对差异表达基因进行验证。通过酵母双杂交筛选与玉米pTAC2互作的蛋白质。【结果】共获得15株T0转基因植株,包括绿色植株（7株）和白色植株（8株）。绿色幼苗中3株为转基因阴性材料,4株为转基因阳性（2株为未编辑,2株为杂合编辑突变）,白色植株(8株)均为转基因阳性纯合编辑。与CK相比,突变体（zmptac2）叶绿体发异常,叶绿素含量显著降低。RNA-seq的结果显示,CK与zmptac2之间共检测到1 367个基因差异表达,其中618个基因上调表达（zmptac2/CK）,749个基因下调表达（zmptac2/CK）。GO富集分析显示,下调基因显著富集到叶绿体和质体中。KEGG分析表明下调表达基因显著富集在苯丙氨酸代谢、酪氨酸代谢和异喹啉生物碱生物合成等途径。选取的15个差异基因表达模式均与测序数据相一致,表明测序结果是可靠的。与CK相比,zmptac2中依赖PEP（plastid-encoded RNA polymerase）转录的基因表达量显著降低,而依赖NEP（nuclear gene-encoded RNA polymerase）转录的基因表达量则显著上升。通过对玉米cDNA文库筛选和互作验证,鉴定出ZmpTAC3与ZmpTAC2存在互作。【结论】ZmpTAC2突变会导致叶绿体早期生物合成受阻,该基因参与叶绿体发育及叶绿素合成,且该种作用是由ZmpTAC2调控PEP相关基因表达而实现的。  相似文献   

3.
The nuclear-encoded light-harvesting chlorophyll a/b-binding proteins(LHCPs) are specifically translocated from the stroma into the thylakoid membrane through the chloroplast signal recognition particle(cp SRP) pathway. The cp SRP is composed of a cp SRP43 protein and a cp SRP54 protein, and it forms a soluble transit complex with LHCP in the chloroplast stroma. Here, we identified the YGL9 gene that is predicted to encode the probable rice cp SRP43 protein from a rice yellow-green leaf mutant. A phylogenetic tree showed that an important conserved protein family, cp SRP43, is present in almost all green photosynthetic organisms such as higher plants and green algae. Sequence analysis showed that YGL9 comprises a chloroplast transit peptide, three chromodomains and four ankyrin repeats, and the chromodomains and ankyrin repeats are probably involved in protein-protein interactions. Subcellular localization showed that YGL9 is localized in the chloroplast. Expression pattern analysis indicated that YGL9 is mainly expressed in green leaf sheaths and leaves. Quantitative real-time PCR analysis showed that the expression levels of genes associated with pigment metabolism, chloroplast development and photosynthesis were distinctly affected in the ygl9 mutant. These results indicated that YGL9 is possibly involved in pigment metabolism, chloroplast development and photosynthesis in rice.  相似文献   

4.
Ininsects,ecdysteroidsaresynthesizedbygenesoftheHalloweenfamilyandplayimportantrolesinseveralkey developmentalevents,includingmoltingandmetamorphosis.However,therolesofthesegenesinAgasicles hygrophila are still largely unknown.In this study, the expression patterns of the two Halloween genesAhCYP307A2andAhCYP314A1weredeterminedbyquantitativePCR(qPCR)atdifferentdevelopmentalstages.Moreover,the functions of these two genes were explored using RNA interference (RNAi), and ovarian development was ob...  相似文献   

5.
Rice is the staple food for about half of the world’s population.Preferred by consumers,aromatic rice is a special type of rice with great commercial value.Cooking and eating qualities and aroma are the major grain qualities favored by most consumers.Currently,most of the available aromatic varieties have low yields and some undesirable agronomic traits.Thus,there is an urgent need to develop better aromatic rice varieties.This work aims to identify rice germplasm lines that have good grain qual...  相似文献   

6.
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7.
Chloroplasts are essential for plant photosynthesis and growth. Many genes have been identified that regulate plant chloroplast development. However, it is not known at a molecular level how these genes regulate chloroplast biogenesis. In this study, we isolated a mutant ygl2(yellow-green leaf2) that exhibited a pigment-defective phenotype. YGL2 encodes a geranylgeranyl reductase, and in mutant ygl2, there was a single base change(T1361 G) located in the third exon of YGL2 that resulted in a missense mutation(L454 R) in the encoded product. Transmission electron microscopy revealed that chloroplast development was impaired in the ygl2 mutant. The expression levels of plastid-encoded genes were significantly altered in the ygl2 mutant. Furthermore, in a yeast two-hybrid assay, we found that YGL2 interacted with the RNA editing factor MORF8.  相似文献   

8.
The rice cultivars carrying dep1 (dense and erect panicle 1) have the potential to achieve both high grain yield and high nitrogen use efficiency (NUE).  However, few studies have focused on the agronomic and physiological performance of those cultivars associated with high yield and high NUE under field conditions.  Therefore, we evaluated the yield performance and NUE of two near-isogenic lines (NILs) carrying DEP1 (NIL-DEP1) and dep1-1 (NIL-dep1) genes under the Nanjing 6 background at 0 and 120 kg N ha–1.  Grain yield and NUE for grain production (NUEg) were 25.5 and 21.9% higher in NIL-dep1 compared to NIL-DEP1 averaged across N treatments and planting years, respectively.  The yield advantage of NIL-dep1 over NIL-DEP1 was mainly due to larger sink size (i.e., higher total spikelet number), grain-filling percentage, total dry matter production, and harvest index.  N utilization rather than N uptake contributed to the high yield of NIL-dep1.  Significantly higher NUEg in NIL-dep1 was associated with higher N and dry matter translocation efficiency, lower leaf and stem N concentration at maturity, and higher glutamine synthetase (GS) activity in leaves.  In conclusion, dep1 improved grain yield and NUE by increasing N and dry matter transport due to higher leaf GS activity under field conditions during the grain-filling period.  相似文献   

9.
Drought and salt stresses, the major environmental abiotic stresses in agriculture worldwide, affect plant growth, crop productivity, and quality. Therefore, developing crops with higher drought and salt tolerance is highly desirable. This study reported the isolation, biological function, and molecular characterization of a novel maspardin gene, OsMas1, from rice. The OsMas1 protein was localized to the cytoplasm. The expression levels of OsMas1 were up-regulated under mannitol, PEG6000, NaCl, ...  相似文献   

10.
Brassica napus is an important cash crop broadly grown for the vegetable and oil values.  Yellow-seeded B. napus is preferred by breeders due to its improved oil and protein quality, less pigments and lignin compared with the black-seeded counterpart.  This study compared the differences in flavonoid and fatty acid contents between yellow rapeseed from the progenies of B. napus–Sinapis alba somatic hybrids and the black-seeded counterpart using RNA-seq analysis.  Through HPLC-PDA-ESI(−)/MS2 analysis, it was found that phenylpropanoids and flavonoids (i.e., isorhamnetin, epicatechin, kaempferol, and other derivatives) in yellow seed were significantly lower than those in black seed.  The fatty acid (FA) content in yellow rapeseed was higher than that in black rapeseed due to the variation of C16:0, C18:0, C18:1, C18:2, and C18:3 contents.  RNA-seq analysis of seeds at four and five weeks after flowering (WAF) indicated that differentially expressed genes (DEGs) between black and yellow rapeseeds were enriched in flavonoid and FA biosynthesis, including BnTT3, BnTT4, BnTT18, and BnFAD2.  Also, genes related to FA biosynthesis, desaturation and elongation (FAD3, LEC1, FUS3, and LPAT2) in yellow seed were up-regulated compared to those in black seed, while genes involved in beta-oxidation cycle (AIM1 and KAT2) of yellow seed were down-regulated compared to those in black seed.  The DEGs related to the variation of flavonoids, phenylpropanoids, and FAs would help improve the knowledge of yellow seed character in B. napus and promote rapeseed improvement.  相似文献   

11.
The fungal pathogen Setosphaeria turcica causes northern corn leaf blight(NCLB), which leads to considerable crop losses. Setosphaeria turcica elaborates a specialized infection structures called appressorium for maize infection.Previously, we demonstrated that the S. turcica triggers an S-phase checkpoint and ATR(Ataxia Telangiectasia and Rad3 related)-dependent self-protective response to DNA genotoxic insults during maize infection. However, how the regulatory mechanism works was still largel...  相似文献   

12.
Arabinogalactan proteins (AGPs) are widely distributed in the plant kingdom and play a vital role during the process of plant sexual reproduction.  In this study, we performed a comprehensive identification of the PbrAGPs expressed in pear pollen and further explored their influences on pollen tube growth.  Among the 187 PbrAGPs that were found to be expressed in pear pollen tubes, 38 PbrAGPs were specifically expressed in pollen according to the RNA-seq data.  The PbrAGPs were divided into two groups of highly expressed and specifically expressed in pear pollen.  We further tested their expression patterns using RT-PCR and RT-qPCR.  Most of the PbrAGPs were expressed in multiple tissues and their expression levels were consistent with reads per kilobase per million map reads (RPKM) values during pollen tube growth, implying that PbrAGPs might be involved in the regulation of pear pollen tube growth.  We also constructed phylogenetic trees to identify the functional genes in pear pollen tube growth.  Therefore, 19 PbrAGPs (PbrAGP1 to PbrAGP19) were selected to test their influences on pollen tube growth.  Recombinant proteins of the 19 PbrAGP-His were purified and used to treat pear pollen, and 11 of the PbrAGP-His recombinant proteins could promote pear pollen tube growth.  Additionally, pollen tube growth was inhibited when the expression levels of PbrAGP1 and PbrAGP5 were knocked down using an antisense oligonucleotide assay.  PbrAGP1 and PbrAGP5 were localized in the plasma membrane and might not alter the distribution of pectin in the pollen tube.  In summary, this study identified the PbrAGPs expressed in pear pollen and lays the foundation for further exploring their functions in pollen tube growth.  相似文献   

13.
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14.
Sugar content is a determinant of apple(Malus×domestica Borkh.) sweetness. However, the molecular mechanism underlying sucrose accumulation in apple fruit remains elusive. Herein, this study reported the role of the sucrose transporter MdSUT2.1 in the regulation of sucrose accumulation in apples. The MdSUT2.1 gene encoded a protein with 612 amino acid residues that could be localized at the plasma membrane when expressed in tobacco leaf protoplasts.MdSUT2.1 was highly expressed in fruit and was ...  相似文献   

15.
Soil microorganisms play important roles in nitrogen transformation.  The aim of this study was to characterize changes in the activity of nitrogen transformation enzymes and the abundance of nitrogen function genes in rhizosphere soil aerated using three different methods (continuous flooding (CF), continuous flooding and aeration (CFA), and alternate wetting and drying (AWD)).  The abundances of amoA ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), nirS, nirK, and nifH genes, and the activities of urease, protease, ammonia oxidase, nitrate reductase, and nitrite reductase were measured at the tillering (S1), heading (S2), and ripening (S3) stages.  We analyzed the relationships of the aforementioned microbial activity indices, in addition to soil microbial biomass carbon (MBC) and soil microbial biomass nitrogen (MBN), with the concentration of soil nitrate and ammonium nitrogen.  The abundance of nitrogen function genes and the activities of nitrogen invertase in rice rhizosphere soil were higher at S2 compared with S1 and S3 in all treatments.  AWD and CFA increased the abundance of amoA and nifH genes, and the activities of urease, protease, and ammonia oxidase, and decreased the abundance of nirS and nirK genes and the activities of nitrate reductase and nitrite reductase, with the effect of AWD being particularly strong.  During the entire growth period, the mean abundances of the AOA amoA, AOB amoA, and nifH genes were 2.9, 5.8, and 3.0 higher in the AWD treatment than in the CF treatment, respectively, and the activities of urease, protease, and ammonia oxidase were 1.1, 0.5, and 0.7 higher in the AWD treatment than in the CF treatment, respectively.  The abundances of the nirS and nirK genes, and the activities of nitrate reductase and nitrite reductase were 73.6, 84.8, 10.3 and 36.5% lower in the AWD treatment than in the CF treatment, respectively.  The abundances of the AOA amoA, AOB amoA, and nifH genes were significantly and positively correlated with the activities of urease, protease, and ammonia oxidase, and the abundances of the nirS and nirK genes were significantly positively correlated with the activities of nitrate reductase.  All the above indicators were positively correlated with soil MBC and MBN.  In sum, microbial activity related to nitrogen transformation in rice rhizosphere soil was highest at S2.  Aeration can effectively increase the activity of most nitrogen-converting microorganisms and MBN, and thus promote soil nitrogen transformation.   相似文献   

16.
Salicylic acid (SA) is an effective elicitor to promote plant defenses and growth.This study aimed to investigate rice(Oryza sativa L.) cv.Khao Dawk Mali 105 treated with salicylic acid (SA)-Ricemate as an enhanced plant protection mechanism against bacterial leaf blight (BLB) disease caused by Xanthomonas oryzae pv.oryzae (Xoo).Results indicated that the use of SA-Ricemate as a foliar spray at concentrations of more than 100 mg L–1 can reduce the severity of BLB disease by 71%.SA-Ric...  相似文献   

17.
The mechanisms that regulate the specificity and maintenance of chicken muscle fiber types remain largely unknown. In mammals, CSRP3 has been shown to play a vital role in the maintenance of typical muscle structure and function. This study investigated the role that CSRP3 plays in chicken skeletal muscle. First, the antibody against chicken CSRP3 protein was prepared, and the expression levels of the mRNA and protein of the CSRP3 gene in four chicken skeletal muscles with different myofiber com...  相似文献   

18.
【目的】 为了丰富和加深人们对植物叶色形成的分子机理认识,对水稻黄绿叶突变体ygl3（yellow green leaf 3）进行表型鉴定和基因克隆,阐明YGL3的分子功能,为解析YGL3调控水稻叶色形成的分子机理奠定基础。【方法】 从水稻中花11 CRISPR-Cas9敲除突变体库中鉴定出2份稳定遗传的等位黄绿叶突变体,命名为ygl3-1与ygl3-2,对突变体的表型进行鉴定,测定野生型和突变体苗期的叶绿素含量,运用透射电镜观察野生型和突变体ygl3的叶绿体结构。利用实时荧光定量PCR分析YGL3的组织表达模式,并使用BioXM 2.6软件对YGL3及其同源蛋白序列进行比对,采用酵母双杂交方法筛选YGL3的互作蛋白。【结果】 在苗期,与野生型相比,突变体ygl3叶片黄化,叶绿素、类胡萝卜素和总光合色素含量显著降低。透射电镜结果表明,突变体ygl3叶绿体形态异常,类囊体片层结构较少,而野生型叶绿体形态正常,类囊体片层结构排列有序。CRISPR-Cas9敲除位点鉴定结果表明,LOC_Os01g73450发生单碱基插入,导致蛋白翻译提前终止,该基因编码351个氨基酸的蛋白突变为55个氨基酸的截短蛋白。与野生型相比,LOC_Os01g73450的表达水平在突变体中显著下调。qRT-PCR结果表明YGL3在水稻根、穗、种子、叶鞘以及叶片中均有表达,且叶片中表达水平最高。YGL3编码一个质体定位的尿嘧啶核苷酸激酶。蛋白氨基酸序列比对表明YGL3蛋白在玉米、高粱、拟南芥等物种中均较为保守,与拟南芥同源蛋白氨基酸的同源性为59.4%。qRT-PCR结果表明,叶绿素合成基因（如HEMC、HEME和URO-D）在突变体ygl3中显著下调,而HEMB、HEMF和HEML等叶绿素合成基因在野生型与突变体之间无显著差异。酵母双杂交系统筛选水稻叶片酵母cDNA文库,发现YGL3与RNA编辑因子MORF8存在互作。【结论】 水稻黄绿叶突变体ygl3的表型是由LOC_Os01g73450突变导致,该基因与已报道的水稻黄绿叶基因YL2/YGL8等位。YGL3在叶片中高度表达,同时YGL3与MORF8在酵母中互作。  相似文献   

19.
Exploring novel high molecular weight glutenin subunits (HMW-GSs) from wild related species is a strategy to improve wheat processing quality.  The objective of the present investigation was to identify the chromosomes of the wheat-alien introgression line N124, derived from the hybridization between Triticum aestivum with Aegilops kotschyi, and characterize the effects on quality-related traits.  Fluorescence in situ hybridization karyotypes showed that N124 is a disomic 1Uk(1A) substitution line.  Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase high-performance liquid chromatography verified N124 expressed two HMW-GSs of the Ae. kotschyi parent.  PacBio RNA sequencing and phylogenetic analysis confirmed that the two HMW-GSs were Ukx and Uky.  Compared to the wheat parent, the substitution line had no obvious agronomic defects except fewer grains per spike but improved several major quality parameters.  It can be served as a donor or bridge material for wheat quality improvement.  相似文献   

20.
Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.  Volatiles emitted from A. annua attract A. lucorum.  Volatile artemisinic acid of A. annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field.  However, little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A. lucorum.  In this study, we collected volatiles from A. annua at the seedling stage by using headspace solid phase microextraction (HS-SPME).  Gas chromatography-mass spectrometry (GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng h–1 artemisinic acid were detected in volatile samples and milled samples, respectively.  Subsequently, a key gene for artemisinic acid synthesis, the cytochrome P450 gene cyp71av1, was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid.  After the addition of exogenous artemisinic alcohol or artemisinic aldehyde, artemisinic acid was identified as the product of the expressed gene.  In electroantennogram (EAG) recordings, 3-day-old adult A. lucorum showed significant electrophysiological responses to artemisinic alcohol, artemisinic aldehyde and artemisinic acid.  Furthermore, 3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L–1, whereas 3-day-old male bugs were attracted significantly by 10 mmol L–1 artemisinic acid and artemisinic aldehyde.  We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A. lucorum.  相似文献   

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