scholarly journals Overexpression of Sesame Polyketide Synthase A Leads to Abnormal Pollen Development in Arabidopsis

2021 ◽  
Author(s):  
Tianyu Li ◽  
Ting Zhou ◽  
Yuanxiao Yang ◽  
Hongyan Liu ◽  
Fang Zhou ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Hybrid Breeding ◽  
Pollen Wall ◽  
Wall Structure ◽  
Ultrastructural Observation ◽  
Wild Type ◽  
In Planta ◽  
Male Sterile ◽  
Crop Species ◽  
Cytoplasmic Inclusions

Abstract Background: Sesame is a great reservoir of bioactive constituents and unique antioxidant components and is widely used for its nutritional and medicinal value. The expanding demands for sesame seeds are putting pressure on sesame breeders to develop reliable high-yielding varieties. Heterosis utilization is an efficient way to increase sesame yield. Polyketide synthases (PKSs) are critical enzymes in the biosynthesis of sporopollenin, a primary component of pollen exine. Their in planta functions are being investigated for application in crop breeding.Results: In this study, we cloned the sesame POLYKETIDE SYNTHASE A (SiPKSA) and examined its function in male sterility. SiPKSA was specifically expressed in sesame flower buds, and its expression was significantly higher in sterile sesame anthers than in fertile anthers at the tetrad and microspore development stage. Further overexpression of SiPKSA in Arabidopsis caused transgenic plants male sterile. Ultrastructural observation showed that the pollen grains of SiPKSA-overexpressing plants contained few cytoplasmic inclusions and exhibited an abnormal pollen wall structure, with a thicker exine layer compared with wild type. In agreement with it, the expression of a set of sporopollenin biosynthesis-related genes and the contents of fatty acids and phenolics were significantly altered in anthers of SiPKSA-overexpressing plants compared with wild type during anther development. Conclusion: These findings highlighted that overexpression of SiPKSA in Arabidopsis might cause excessive sporopollenin biosynthesis to influence pollen and pollen wall development, leading to male sterile, suggesting that its manipulation might improve hybrid breeding in sesame and other crop species.

scholarly journals Tetraketide α-pyrone reductases in sporopollenin synthesis pathway in Gerbera hybrida: diversification of the minor function

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Lingping Zhu ◽  
Teng Zhang ◽  
Teemu H. Teeri
Keyword(s):  
Pollen Development ◽  
Polyketide Synthase ◽  
Land Plant ◽  
Vital Role ◽  
Pollen Wall ◽  
Major Constituent ◽  
Minor Role ◽  
Male Sterile ◽  
Gerbera Hybrida ◽  
Secondary Role

AbstractThe structurally robust biopolymer sporopollenin is the major constituent of the exine layer of pollen wall and plays a vital role in plant reproductive success. The sporopollenin precursors are synthesized through an ancient polyketide biosynthetic pathway consisting of a series of anther-specific enzymes that are widely present in all land plant lineages. Tetraketide α-pyrone reductase 1 (TKPR1) and TKPR2 are two reductases catalyzing the final reduction of the carbonyl group of the polyketide synthase-synthesized tetraketide intermediates to hydroxylated α-pyrone compounds, important precursors of sporopollenin. In contrast to the functional conservation of many sporopollenin biosynthesis associated genes confirmed in diverse plant species, TKPR2’s role has been addressed only in Arabidopsis, where it plays a minor role in sporopollenin biosynthesis. We identified in gerbera two non-anther-specific orthologues of AtTKPR2, Gerbera reductase 1 (GRED1) and GRED2. Their dramatically expanded expression pattern implies involvement in pathways outside of the sporopollenin pathway. In this study, we show that GRED1 and GRED2 are still involved in sporopollenin biosynthesis with a similar secondary role as AtTKPR2 in Arabidopsis. We further show that this secondary role does not relate to the promoter of the gene, AtTKPR2 cannot rescue pollen development in Arabidopsis even when controlled by the AtTKPR1 promoter. We also identified the gerbera orthologue of AtTKPR1, GTKPR1, and characterized its crucial role in gerbera pollen development. GTKPR1 is the predominant TKPR in gerbera pollen wall formation, in contrast to the minor roles GRED1 and GRED2. GTKPR1 is in fact an excellent target for engineering male-sterile gerbera cultivars in horticultural plant breeding.

scholarly journals OsMS188 Is a Key Regulator of Tapetum Development and Sporopollenin Synthesis in Rice

Rice ◽  
2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yu Han ◽  
Si-Da Zhou ◽  
Jiong-Jiong Fan ◽  
Lei Zhou ◽  
Qiang-Sheng Shi ◽  
...  
Keyword(s):  
Pollen Wall ◽  
Myb Transcription Factor ◽  
Male Sterile ◽  
Mobility Shift ◽  
Crop Species ◽  
Wall Formation ◽  
Biochemical Assays ◽  
Anther Cuticle ◽  
Electrophoretic Mobility Shift Assays ◽  
Downstream Gene Expression

Abstract Background During anther development, the tapetum provides essential nutrients and materials for pollen development. In rice, multiple transcription factors and enzymes essential for tapetum development and pollen wall formation have been cloned from male-sterile lines. Results In this study, we obtained several lines in which the MYB transcription factor OsMS188 was knocked out through the CRISPR-Cas9 approach. The osms188 lines exhibited a male-sterile phenotype with aberrant development and degeneration of tapetal cells, absence of the sexine layer and defective anther cuticles. CYP703A3, CYP704B2, OsPKS1, OsPKS2, DPW and ABCG15 are sporopollenin synthesis and transport-related genes in rice. Plants with mutations in these genes are male sterile, with a defective sexine layer and anther cuticle. Further biochemical assays demonstrated that OsMS188 binds directly to the promoters of these genes to regulate their expression. UDT1, OsTDF1, TDR, bHLH142 and EAT1 are upstream regulators of rice tapetum development. Electrophoretic mobility shift assays (EMSAs) and activation assays revealed that TDR directly regulates OsMS188 expression. Additionally, protein interaction assays indicated that TDR interacts with OsMS188 to regulate downstream gene expression. Conclusion Overall, OsMS188 is a key regulator of tapetum development and pollen wall formation. The gene regulatory network established in this work may facilitate future investigations of fertility regulation in rice and in other crop species.

scholarly journals Generating Novel Male Sterile Tomatoes by Editing Respiratory Burst Oxidase Homolog Genes

2022 ◽  
Vol 12 ◽  
Author(s):  
Xiaojuan Dai ◽  
Huanan Han ◽  
Wei Huang ◽  
Lianghui Zhao ◽  
Minglei Song ◽  
...  
Keyword(s):  
Respiratory Burst ◽  
Hybrid Breeding ◽  
Wild Type ◽  
Male Sterile ◽  
Ros Metabolism ◽  
Reproduction Process ◽  
Self Fertilization ◽  
Mutant Lines ◽  
Respiratory Burst Oxidase

Hybrid breeding of tomatoes (Solanum lycopersicum), an important vegetable crop, is an effective way to improve yield and enhance disease and stress resistance. However, the efficiency of tomato hybridization is hindered by self-fertilization, which can be overcome using male sterile lines. It has been reported that reactive oxygen species (ROS) act as a key regulator for anther development, mediated by RBOH (Respiratory Burst Oxidase Homolog) genes. Here, two tomato anther-expressed genes, LeRBOH (Solyc01g099620) and LeRBOHE (Solyc07g042460), were selected to cultivate novel tomato male sterile strains. By using a CRISPR/Cas9 system with a two-sgRNA module, the lerboh, lerbohe, and lerboh lerbohe mutant lines were generated, among which the lerbohe and lerboh lerbohe mutants displayed complete male sterility but could accept wild-type pollens and produce fruits normally. Further analysis uncovered significantly decreased ROS levels and abnormal programmed cell death in lerboh lerbohe anthers, indicating a key role of ROS metabolism in tomato pollen development. Taken together, our work demonstrates a successful application of gene editing via CRISPR/Cas9 in generating male sterile tomatoes and afforded helpful information for understanding how RBOH genes regulating tomato reproduction process.

scholarly journals A single nucleotide polymorphism in an R2R3 MYB transcription factor gene triggers the male sterility in soybean ms6 (Ames1)

2021 ◽  
Author(s):  
Junping Yu ◽  
Guolong Zhao ◽  
Wei Li ◽  
Ying Zhang ◽  
Peng Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Glycine Max ◽  
Male Sterility ◽  
Bulked Segregant Analysis ◽  
Soybean Protein ◽  
Anther Development ◽  
Hybrid Breeding ◽  
Myb Transcription Factor ◽  
Male Sterile ◽  
R2r3 Myb

Abstract Key message Identification and functional analysis of the male sterile gene MS6 in Glycine max. Abstract Soybean (Glycine max (L.) Merr.) is an important crop providing vegetable oil and protein. The male sterility-based hybrid breeding is a promising method for improving soybean yield to meet the globally growing demand. In this research, we identified a soybean genic male sterile locus, MS6, by combining the bulked segregant analysis sequencing method and the map-based cloning technology. MS6, highly expressed in anther, encodes an R2R3 MYB transcription factor (GmTDF1-1) that is homologous to Tapetal Development and Function 1, a key factor for anther development in Arabidopsis and rice. In male sterile ms6 (Ames1), the mutant allele contains a missense mutation, leading to the 76th leucine substituted by histidine in the DNA binding domain of GmTDF1-1. The expression of soybean MS6 under the control of the AtTDF1 promoter could rescue the male sterility of attdf1 but ms6 could not. Additionally, ms6 overexpression in wild-type Arabidopsis did not affect anther development. These results evidence that GmTDF1-1 is a functional TDF1 homolog and L76H disrupts its function. Notably, GmTDF1-1 shows 92% sequence identity with another soybean protein termed as GmTDF1-2, whose active expression also restored the fertility of attdf1. However, GmTDF1-2 is constitutively expressed at a very low level in soybean, and therefore, not able to compensate for the MS6 deficiency. Analysis of the TDF1-involved anther development regulatory pathway showed that expressions of the genes downstream of TDF1 are significantly suppressed in ms6, unveiling that GmTDF1-1 is a core transcription factor regulating soybean anther development.

scholarly journals The PKS4 Gene of Fusarium graminearum Is Essential for Zearalenone Production

2006 ◽  
Vol 72 (6) ◽  
pp. 3924-3932 ◽  
Author(s):  
Erik Lys�e ◽  
Sonja S. Klemsdal ◽  
Karen R. Bone ◽  
Rasmus J. N. Frandsen ◽  
Thomas Johansen ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
High Performance ◽  
Polyketide Synthase ◽  
Wild Type ◽  
Root Infection ◽  
Enoyl Reductase ◽  
Transformation Protocol ◽  
Chromatography Analysis ◽  
In The Wild ◽  
High Level

ABSTRACT Zearalenones are produced by several Fusarium species and can cause reproductive problems in animals. Some aurofusarin mutants of Fusarium pseudograminearum produce elevated levels of zearalenone (ZON), one of the estrogenic mycotoxins comprising the zearalenones. An analysis of transcripts from polyketide synthase genes identified in the Fusarium graminearum database was carried out for these mutants. PKS4 was the only gene with an enoyl reductase domain that had a higher level of transcription in the aurofusarin mutants than in the wild type. An Agrobacterium tumefaciens-mediated transformation protocol was used to replace the central part of the PKS4 gene with a hygB resistance gene through double homologous recombination in an F. graminearum strain producing a high level of ZON. PCR and Southern analysis of transformants were used to identify isolates with single insertional replacements of PKS4. High-performance liquid chromatography analysis showed that the PKS4 replacement mutant did not produce ZON. Thus, PKS4 encodes an enzyme required for the production of ZON in F. graminearum. Barley root infection studies revealed no alteration in the pathogenicity of the PKS4 mutant compared to the pathogenicity of the wild type. The expression of PKS13, which is located in the same cluster as PKS4, decreased dramatically in the mutant, while transcription of PKS4 was unchanged. This differential expression may indicate that ZON or its derivatives do not regulate expression of PKS4 and that the PKS4-encoded protein or its product stimulates expression of PKS13. Furthermore, both the lack of aurofusarin and ZON influenced the expression of other polyketide synthases, demonstrating that one polyketide can influence the expression of others.

Isolation of a chitinase overproducing mutant of Streptomyces peucetius defective in daunorubicin biosynthesis

2000 ◽  
Vol 46 (10) ◽  
pp. 956-960 ◽  
Author(s):  
Kuzhandhaivel S Vetrivel ◽  
Kuppamuthu Dharmalingam
Keyword(s):  
Biosynthetic Pathway ◽  
Time Course ◽  
Polyketide Synthase ◽  
Protein Profile ◽  
Restriction Enzymes ◽  
Wild Type ◽  
Anthracycline Antibiotic ◽  
Streptomyces Peucetius ◽  
Chitinase Production ◽  
Major Alteration

Streptomyces peucetius, producer of the antitumor anthracycline antibiotic daunorubicin, was mutagenized, and mutants defective in daunorubicin biosynthesis were screened. One mutant (SPVI), which failed to produce daunorubicin, was found to overproduce an extracellular chitinase. Time course analyses of chitinase production and of the extracellular protein profile showed that the increase in activity is due to increased synthesis of the enzyme protein. The production of chitinase in SPVI was repressed by glucose as in the case of wild-type S. peucetius. PFGE analysis of VspI restriction fragments of S. peucetius and SPVI showed that there was no major alteration in the mutant genome. The hybridization pattern of S. peucetius and SPVI genomic DNA digested with various restriction enzymes was identical when probed with dnrUVJI genes of the S. peucetius daunorubicin cluster and chiA of Streptomyces lividans 66. The possible step affected in the daunorubicin biosynthetic pathway could be a polyketide synthase, since aklanonic acid, the earliest detectable intermediate in the daunorubicin pathway, was not synthesized in SPVI.Key words: Streptomyces peucetius, chitinase, daunorubicin, NTG mutagenesis.

scholarly journals A Rapid Pipeline for Pollen- and Anther-Specific Gene Discovery Based on Transcriptome Profiling Analysis of Maize Tissues

2021 ◽  
Vol 22 (13) ◽  
pp. 6877
Author(s):  
Yannan Shi ◽  
Yao Li ◽  
Yongchao Guo ◽  
Eli James Borrego ◽  
Zhengyi Wei ◽  
...  
Keyword(s):  
Seed Production ◽  
Late Stage ◽  
Mature Pollen ◽  
Hybrid Breeding ◽  
Large Set ◽  
Male Sterile ◽  
Reproductive Process ◽  
Maize Pollen ◽  
Cis Element ◽  
Mature Anther

Recently, crop breeders have widely adopted a new biotechnology-based process, termed Seed Production Technology (SPT), to produce hybrid varieties. The SPT does not produce nuclear male-sterile lines, and instead utilizes transgenic SPT maintainer lines to pollinate male-sterile plants for propagation of nuclear-recessive male-sterile lines. A late-stage pollen-specific promoter is an essential component of the pollen-inactivating cassette used by the SPT maintainers. While a number of plant pollen-specific promoters have been reported so far, their usefulness in SPT has remained limited. To increase the repertoire of pollen-specific promoters for the maize community, we conducted a comprehensive comparative analysis of transcriptome profiles of mature pollen and mature anthers against other tissue types. We found that maize pollen has much less expressed genes (>1 FPKM) than other tissue types, but the pollen grain has a large set of distinct genes, called pollen-specific genes, which are exclusively or much higher (100 folds) expressed in pollen than other tissue types. Utilizing transcript abundance and correlation coefficient analysis, 1215 mature pollen-specific (MPS) genes and 1009 mature anther-specific (MAS) genes were identified in B73 transcriptome. These two gene sets had similar GO term and KEGG pathway enrichment patterns, indicating that their members share similar functions in the maize reproductive process. Of the genes, 623 were shared between the two sets, called mature anther- and pollen-specific (MAPS) genes, which represent the late-stage pollen-specific genes of the maize genome. Functional annotation analysis of MAPS showed that 447 MAPS genes (71.7% of MAPS) belonged to genes encoding pollen allergen protein. Their 2-kb promoters were analyzed for cis-element enrichment and six well-known pollen-specific cis-elements (AGAAA, TCCACCA, TGTGGTT, [TA]AAAG, AAATGA, and TTTCT) were found highly enriched in the promoters of MAPS. Interestingly, JA-responsive cis-element GCC box (GCCGCC) and ABA-responsive cis-element-coupling element1 (ABRE-CE1, CCACC) were also found enriched in the MAPS promoters, indicating that JA and ABA signaling likely regulate pollen-specific MAPS expression. This study describes a robust and straightforward pipeline to discover pollen-specific promotes from publicly available data while providing maize breeders and the maize industry a number of late-stage (mature) pollen-specific promoters for use in SPT for hybrid breeding and seed production.

Identification of a DNA region required for growth of Pseudomonas syringae pv. tomato on tomato plants

1992 ◽  
Vol 38 (9) ◽  
pp. 883-890 ◽  
Author(s):  
Dennis P. Jackson ◽  
Douglas A. Gray ◽  
Vincent L. Morris ◽  
Diane A. Cuppels
Keyword(s):  
Organic Acids ◽  
Pseudomonas Syringae ◽  
Acid Metabolism ◽  
Carbon Sources ◽  
Hypersensitive Reaction ◽  
Wild Type ◽  
In Planta ◽  
Tomato Plants ◽  
Tartaric Acids ◽  
Nonpathogenic Strain

The prototrophic Pseudomonas syringae pv. tomato mutant DC3481, which is the result of a single-site Tn5 insertion, cannot grow and cause disease on tomato plants and cannot use the major organic acids of tomato, i.e., citric, malic, succinic, and tartaric acids, as sole carbon sources. Although nonpathogenic, strain DC3481 can still induce a hypersensitive reaction in nonhost plants. We have identified a 30-kb fragment of P. syringae pv. tomato wild-type DNA that can complement this mutant. EcoRI fragments from this region were subcloned and individually subjected to functional complementation analysis. The 3.8-kb fragment, which was the site of the Tn5 insertion, restored pathogenicity and the ability to use all the major organic acids of tomato as carbon sources. It shares sequence homology with several P. syringae pathovars but not other bacterial tomato pathogens. Our results indicate that sequences on the 3.8-kb EcoRI fragment are required for both the ability to grow on tomato leaves (and thus cause disease) and the utilization of carboxylic acids common to tomato. The 3.8-kb fragment may contain a sequence (or sequences) that regulates both traits. Key words: Pseudomonas syringae pv. tomato, phytopathogenicity, Tn5, tricarboxylic acid metabolism, bacterial speck, growth in planta.

scholarly journals Extension of thein vivohaploid induction system from maize to wheat

2019 ◽  
Author(s):  
Chenxu Liu ◽  
Yu Zhong ◽  
Xiaolong Qi ◽  
Ming Chen ◽  
Zongkai Liu ◽  
...  
Keyword(s):  
Pollen Viability ◽  
Haploid Induction ◽  
Male Sterile ◽  
Crop Breeding ◽  
Crop Species ◽  
Ploidy Levels ◽  
New Approach ◽  
Knock Out ◽  
Haploid Production

AbstractDoubled haploid breeding technology has been one of the most important techniques for accelerating crop breeding. In compare toin vivohaploid induction in maize, which is efficient and background independent, wheat haploid production by interspecific hybridization pollinated with maize is influenced by genetic background and requires rescue of young embryos. Here, we analyzed the homologues of maize haploid induction geneMTL/ZmPLA1/NLDin several crop species systematically, the homologues are highly conserved in sorghum, millet and wheat etc. Since wheat is a very important polyploidy crop, as a proof of concept, we demonstrated that thein vivohaploid induction method could be extended from diploid maize to hexaploid wheat by knocking out the wheat homologues (TaPLAs). Result showed that double knock-out mutation could trigger wheat haploid induction at ~ 2%-3%, accompanied by 30% - 60% seed setting rate. The performance of haploid wheat individual showed shorter plant, narrower leaves and male sterile. Our results also revealed that knockout ofTaPLA-A andTaPLA-D do not affect pollen viability. This study not only confirmed the function of the induction gene and explored a new approach for haploid production in wheat, but also provided an example that thein vivohaploid induction could be applied in more crop species with different ploidy levels. Furthermore, by combining with gene editing, it would be a fast and powerful platform for traits improvement in polyploidy crops breeding.

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