scholarly journals Transcriptome and Hormone Comparison of Three Cytoplasmic Male Sterile Systems in Brassica napus

2018 ◽  
Vol 19 (12) ◽  
pp. 4022 ◽  
Author(s):  
Bingli Ding ◽  
Mengyu Hao ◽  
Desheng Mei ◽  
Qamar Zaman ◽  
Shifei Sang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Stress Responses ◽  
Developmental Stages ◽  
Plant Mitochondria ◽  
Research Work ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Tetrad Stage ◽  
Cms Line ◽  
Aba Content

The interaction between plant mitochondria and the nucleus markedly influences stress responses and morphological features, including growth and development. An important example of this interaction is cytoplasmic male sterility (CMS), which results in plants producing non-functional pollen. In current research work, we compared the phenotypic differences in floral buds of different Brassica napus CMS (Polima, Ogura, Nsa) lines with their corresponding maintainer lines. By comparing anther developmental stages between CMS and maintainer lines, we identified that in the Nsa CMS line abnormality occurred at the tetrad stage of pollen development. Phytohormone assays demonstrated that IAA content decreased in sterile lines as compared to maintainer lines, while the total hormone content was increased two-fold in the S2 stage compared with the S1 stage. ABA content was higher in the S1 stage and exhibited a two-fold decreasing trend in S2 stage. Sterile lines however, had increased ABA content at both stages compared with the corresponding maintainer lines. Through transcriptome sequencing, we compared differentially expressed unigenes in sterile and maintainer lines at both (S1 and S2) developmental stages. We also explored the co-expressed genes of the three sterile lines in the two stages and classified these genes by gene function. By analyzing transcriptome data and validating by RT-PCR, it was shown that some transcription factors (TFs) and hormone-related genes were weakly or not expressed in the sterile lines. This research work provides preliminary identification of the pollen abortion stage in Nsa CMS line. Our focus on genes specifically expressed in sterile lines may be useful to understand the regulation of CMS.

scholarly journals Cytological observation of anther structure and genetic investigation of a thermo-sensitive genic male sterile line 373S in Brassica napus L

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Yanyan Sun ◽  
Dongsuo Zhang ◽  
Zhenzhen Wang ◽  
Yuan Guo ◽  
Xiaomin Sun ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Male Sterility ◽  
Spontaneous Mutation ◽  
Hybrid Breeding ◽  
Pcr Analysis ◽  
Cytological Observation ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Genetic Investigation ◽  
Temperature Changes

Abstract Background Photoperiod and/or thermo-sensitive male sterility is an effective pollination control system in crop two-line hybrid breeding. We previously discovered the spontaneous mutation of a partially male sterile plant and developed a thermo-sensitive genic male sterile (TGMS) line 373S in Brassica napus L. The present study characterized this TGMS line through cytological observation, photoperiod/ temperature treatments, and genetic investigation. Results Microscopic observation revealed that the condensed cytoplasm and irregular exine of microspores and the abnormal degradation of tapetum are related to pollen abortion. Different temperature and photoperiod treatments in field and growth cabinet conditions indicated that the fertility alteration of 373S was mainly caused by temperature changes. The effects of photoperiod and interaction between temperature and photoperiod were insignificant. The critical temperature leading to fertility alteration ranged from 10 °C (15 °C/5 °C) to 12 °C (17 °C/7 °C), and the temperature-responding stage was coincident with anther development from pollen mother cell formation to meiosis stages. Genetic analysis indicated that the TGMS trait in 373S was controlled by one pair of genes, with male sterility as the recessive. Multiplex PCR analysis revealed that the cytoplasm of 373S is pol type. Conclusions Our study suggested that the 373S line in B. napus has a novel thermo-sensitive gene Bnmst1 in Pol CMS cytoplasm background, and its fertility alteration is mainly caused by temperature changes. Our results will broaden the TGMS resources and lay the foundation for two-line hybrid breeding in B. napus.

scholarly journals Unraveling the Genetic Basis of Fertility Restoration for Cytoplasmic Male Sterile Line WNJ01A Originated From Brassica juncea in Brassica napus

2021 ◽  
Vol 12 ◽  
Author(s):  
Qian Yang ◽  
Xiaoyi Nong ◽  
Jize Xu ◽  
Fan Huang ◽  
Fang Wang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
26S Proteasome ◽  
Morphological Observation ◽  
Male Sterile ◽  
Bac Clone ◽  
Cell Stage ◽  
Cms Line ◽  
Candidate Interval

Crosses that lead to heterosis have been widely used in the rapeseed (Brassica napus L.) industry. Cytoplasmic male sterility (CMS)/restorer-of-fertility (Rf) systems represent one of the most useful tools for rapeseed production. Several CMS types and their restorer lines have been identified in rapeseed, but there are few studies on the mechanisms underlying fertility restoration. Here, we performed morphological observation, map-based cloning, and transcriptomic analysis of the F2 population developed by crossing the CMS line WNJ01A with its restorer line Hui01. Paraffin-embedded sections showed that the sporogenous cell stage was the critical pollen degeneration period, with major sporogenous cells displaying loose and irregular arrangement in sterile anthers. Most mitochondrial electron transport chain (mtETC) complex genes were upregulated in fertile compared to sterile buds. Using bulked segregant analysis (BSA)-seq to analyze mixed DNA pools from sterile and fertile F2 buds, respectively, we identified a 6.25 Mb candidate interval where Rfw is located. Using map-based cloning experiments combined with bacterial artificial chromosome (BAC) clone sequencing, the candidate interval was reduced to 99.75 kb and two pentatricopeptide repeat (PPR) genes were found among 28 predicted genes in this interval. Transcriptome sequencing showed that there were 1679 DEGs (1023 upregulated and 656 downregulated) in fertile compared to sterile F2 buds. The upregulated differentially expressed genes (DEGs) were enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) lysine degradation pathway and phenylalanine metabolism, and the downregulated DEGs were enriched in cutin, suberine, and wax biosynthesis. Furthermore, 44 DEGs were involved in pollen and anther development, such as tapetum, microspores, and pollen wall development. All of them were upregulated except a few such as POE1 genes (which encode Pollen Ole e I allergen and extensin family proteins). There were 261 specifically expressed DEGs (9 and 252 in sterile and fertile buds, respectively). Regarding the fertile bud-specific upregulated DEGs, the ubiquitin–proteasome pathway was enriched. The top four hub genes in the protein–protein interaction network (BnaA09g56400D, BnaA10g18210D, BnaA10g18220D, and BnaC09g41740D) encode RAD23d proteins, which deliver ubiquitinated substrates to the 26S proteasome. These findings provide evidence on the pathways regulated by Rfw and improve our understanding of fertility restoration.

Sterility and variation resulting from the transfer of polima cytoplasmic male sterility from Brassica napus into Brassica chinensis

1997 ◽  
Vol 128 (3) ◽  
pp. 299-301 ◽  
Author(s):  
B. HU ◽  
F. CHEN ◽  
Q. LI
Keyword(s):  
Control System ◽  
Brassica Napus ◽  
Interspecific Crosses ◽  
Hybrid Seed Production ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Brassica Chinensis ◽  
Full Flowering ◽  
Cms Line ◽  
Pollination Control

Male sterile combinations made from interspecific crosses between the polima CMS line of Brassica napus and varieties of B. chinensis were backcrossed to BC3. Twenty-six selfing lines from B. chinensis were tested for their ability to either maintain complete sterility or to restore fertility in crosses with the polima male sterile line. Results show that four of these hybrids were completely male sterile and two were fertile. The sterility of the B. chinensis with polima cytoplasm was much more stable than male sterile lines with B. campestris and B. chinensis cytoplasm, which were sterile before full flowering but progressively became fertile as flowering proceeded. The results suggest that polima cytoplasm could be a suitable male sterile-inducing allocytoplasm for B. chinensis, as both maintainers and restorers are available, and could supply a reliable pollination control system for hybrid seed production in this species.

scholarly journals Deciphering the dynamic gene expression patterns of pollen abortion in a male sterile line of Avena sativa through transcriptome analysis at different developmental stages

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Lijun Zhang ◽  
Mingchuan Ma ◽  
Lin Cui ◽  
Longlong Liu
Keyword(s):  
Gene Expression ◽  
Male Sterility ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Hybrid Breeding ◽  
Gene Expression Patterns ◽  
Hybrid Seed Production ◽  
Isogenic Line ◽  
Male Sterile ◽  
Pollen Abortion

Abstract Background Male sterility (MS) has important applications in hybrid seed production, and the abortion of anthers has been observed in many plant species. While most studies have focused on the genetic factors affecting male sterility, the dynamic gene expression patterns of pollen abortion in male sterile lines have not been fully elucidated. In addition, there is still no hybrid oat that is commercially planted due to the lack of a suitable system of male sterility for hybrid breeding. Results In this study, we cultivated a male sterile oat line and a near-isogenic line by crossbreeding to elucidate the expression patterns of genes that may be involved in sterility. The first reported CA male sterile (CAMS) oat line was used for cross-testing and hybridization experiments and was confirmed to exhibit a type of nuclear sterility controlled by recessive genes. Oat stamens of two lines were sampled at four different developmental stages separately. Paired-end RNA sequencing was performed for each sample and generated 252.84 Gb sequences. There were 295,462 unigenes annotated in public databases in all samples, and we compared the histological characteristics and transcriptomes of oat stamens from the two oat lines at different developmental stages. Our results demonstrate that the sterility of the male sterile oat line occurs in the early stage of stamen development and is primarily attributable to abnormal meiosis and the excessive accumulation of superoxide. Conclusions To the best of our knowledge, this study is the first to decipher the dynamic expression profiles of pollen abortion CAMS and CA male fertile (CAMF) oat lines, which may represent a valuable resource for further studies attempting to understand pollen abortion and anther development in oats.

Identification of genes related to the regulation of anther and pollen development in Mu-type cytoplasmic male sterile wheat (Triticum aestivum) by transcriptome analysis

2019 ◽  
Vol 70 (4) ◽  
pp. 306 ◽  
Author(s):  
Wei Li ◽  
Zihan Liu ◽  
Chang Meng ◽  
Yulin Jia ◽  
Lingli Zhang ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Wheat Breeding ◽  
Abnormal Development ◽  
Nutrient Deficiencies ◽  
Hybrid Wheat ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Pectin Degradation ◽  
Cytoplasmic Male Sterile ◽  
Cms Line

Cytoplasmic male sterile (CMS) lines are important tools for hybrid production but they cannot produce viable pollen. Breeding new CMS lines and studying their sterility mechanism in wheat (Triticum aestivum L.) greatly facilitates the process of hybrid wheat breeding. We conducted transcriptome sequencing for a recently identified Mu-CMS line with Aegilops uniaristata Vis. cytoplasm, named U706A, and its isonuclear maintainer line (706B) at the binucleate stage, which was a critical period when abortion occurred. We found that most of the genes involved in phosphatidylinositol metabolism and pectin degradation were downregulated, as well as genes encoding the MYB21 and MYC2 transcription factors, in U706A compared with 706B. In addition, pectin contents indicated that the production of pectin has been enhanced from the binucleate stage to the trinucleate stage, owing to the downregulation of pectin-degradation-related genes in U706A at the binucleate stage, which confirmed the reliability of the sequencing results. We also discovered that the accumulation period of pectin content in U706A is abnormal compared with 706B, which may be an important reason for abortion. Some differentially expressed genes that might be related to the sterile phenotype were verified by quantitative RT-PCR. Therefore, we suggest that the downregulation of these genes possibly leads to the anther not to crack; the tapetum and microspore membrane system is less metabolised, and the abnormal pectin accumulation results in microspore nutrient deficiencies and abnormal development. These findings provide novel insights into the mechanism responsible for pollen abortion in CMS, which may facilitate hybrid wheat breeding.

scholarly journals Integrated cyto-physiological and proteomic analyses reveal new insight into CMS mechanism in a novel upland cotton CMS line LD6A

2022 ◽  
Author(s):  
Zheng Jie ◽  
Aziz Khan ◽  
Zhou Bujin ◽  
Zhou Qiong ◽  
Najeeb Ullah ◽  
...  
Keyword(s):  
Fertility Restoration ◽  
Hybrid Vigor ◽  
Male Sterile ◽  
Maintainer Line ◽  
Tetrad Stage ◽  
Significant Differential Expression ◽  
Mitochondrial Ribosome ◽  
Cms Line ◽  
Analytical Approaches ◽  
Insight Into

Cytoplasmic male sterile (CMS) system has extensively been exploited for hybrid vigor in plant breeding programs. However, its application in many crops is limited due to poor understanding of molecular mechanism of fertility restoration. Using advanced analytical approaches, we elucidated molecular pathways regulating CMS induction and fertility restoration in cotton. Reproductive structures of a novel CMS (LD6A) and its maintainer (LD6B) line were analyzed for physiological and proteomic changes during the development process. Significant differential expression of proteins, such as Abrin, malate dehydrogenase, malic enzyme, isocitrate dehydrogenase, histone acetyltransferase was observed in CMS and its maintainer line. Transmission electron micrographs of anther tapetum showed that inner ridge of CMS mitochondria was relatively indistinct than that of LD6B with narrower membranous space at tetrad stage. Further, relatively higher reactive oxygen species were accumulated in the anther of CMS than its maintainer line at pollen mother cell and tetrad stage. We suggest that abnormal sequence of mitochondrial ribosome gene rps4 and rpl10 and high expression of ribosome-inactivating protein gene Abrin in CMS line damaged mitochondrial membrane and consequently induced pollen sterility. These data provide new insight into CMS mechanism in cotton crops and a tool to develop new CMS germplasm resources.

scholarly journals Comparative Transcriptome Analysis Reveals the Cause for Accumulation of Reactive Oxygen Species During Pollen Abortion in Cytoplasmic Male-Sterile Kenaf Line 722HA

2019 ◽  
Vol 20 (21) ◽  
pp. 5515
Author(s):  
Zhou ◽  
Liu ◽  
Chen ◽  
Liu ◽  
Wang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Molecular Mechanism ◽  
Reactive Oxygen ◽  
Hybrid Breeding ◽  
Oxygen Species ◽  
Ros Scavenging ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Cms Line ◽  
Recurrent Backcrossing

Cytoplasmic male sterility (CMS) is a maternally inherited trait used for hybrid production in plants, a novel kenaf CMS line 722HA was derived from the thermo-sensitive male-sterile mutant ‘HMS’ by recurrent backcrossing with 722HB. The line 722HA has great potential for hybrid breeding in kenaf. However, the underlying molecular mechanism that controls pollen abortion in 722HA remains unclear, thus limiting the full utilization of this line. To understand the possible mechanism governing pollen abortion in 722HA, cytological, transcriptomic, and biochemical analyses were carried out to compare the CMS line 722HA and its maintainer line 722HB. Cytological observations of the microspore development revealed premature degradation of the tapetum at the mononuclear stage, which resulted in pollen dysfunction. The k-means clustering analysis of differentially expressed genes (DEGs) revealed that these genes are related to processes associated with the accumulation of reactive oxygen species (ROS), including electron transport chain, F1F0-ATPase proton transport, positive regulation of superoxide dismutase (SOD), hydrogen peroxide catabolic, and oxidation-reduction. Biochemical analysis indicated that ROS-scavenging capability was lower in 722HA than in 722HB, resulting in an accumulation of excess ROS, which is consistent with the transcriptome results. Taken together, these results demonstrate that excessive ROS accumulation may affect the normal development of microspores. Our study provides new insight into the molecular mechanism of pollen abortion in 722HA and will promote further studies of kenaf hybrids.

scholarly journals Differentially Expressed Genes between Carrot Petaloid Cytoplasmic Male Sterile and Maintainer during Floral Development

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Bo Liu ◽  
Chenggang Ou ◽  
Shumin Chen ◽  
Qiongwen Cao ◽  
Zhiwei Zhao ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Floral Development ◽  
Developmental Stages ◽  
Nuclear Interaction ◽  
Differentially Expressed ◽  
Intrinsic Stress ◽  
Mads Box ◽  
Mads Box Genes ◽  
Male Sterile ◽  
Cms Line

AbstractPetaloid cytoplasmic male sterility (CMS) is a maternally inherited loss of male fertility due to the complete conversion of stamens into petal-like organs, and CMS lines have been widely utilized in carrot breeding. Petaloid CMS is an ideal model not only for studying the mitochondrial–nuclear interaction but also for discovering genes that are essential for floral organ development. To investigate the comprehensive mechanism of CMS and homeotic organ alternation during carrot flower development, we conducted transcriptome analysis between the petaloid CMS line (P2S) and its maintainer line (P2M) at four flower developmental stages (T1–T4). A total of 2838 genes were found to be differentially expressed, among which 1495 genes were significantly downregulated and 1343 genes were significantly upregulated in the CMS line. Functional analysis showed that most of the differentially expressed genes (DEGs) were involved in protein processing in the endoplasmic reticulum, plant hormone signal transduction, and biosynthesis. A total of 16 MADS-box genes were grouped into class A, B, C, and E, but not class D, genes. Several key genes associated with oxidative phosphorylation showed continuously low expression from stage T2 in P2S, and the expression of DcPI and DcAG-like genes also greatly decreased at stage T2 in P2S. This indicated that energy deficiency might inhibit the expression of B- and C-class MADS-box genes resulting in the conversion of stamens into petals. Stamen petaloidy may act as an intrinsic stress, upregulating the expression of heat shock protein (HSP) genes and MADS-box genes at stages T3 and T4 in P2S, which results in some fertile revertants. This study will provide a better understanding of carrot petaloid CMS and floral development as a basis for further research.

Understanding male sterility in Miconia species (Melastomataceae): a morphological approach

2012 ◽  
Vol 60 (6) ◽  
pp. 506 ◽  
Author(s):  
Priscila Andressa Cortez ◽  
Sandra Maria Carmello-Guerreiro ◽  
Simone Pádua Teixeira
Keyword(s):  
Pollen Tube ◽  
Male Sterility ◽  
Developmental Stages ◽  
Rare Event ◽  
Pollen Grains ◽  
Pollen Grain ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Large Numbers ◽  
Morphological Approach

Pollen abortion occurs in virtually all species and often does not prejudice reproductive success. However, large numbers of abnormal pollen grains are characteristic of some groups. Among them is Miconia, in which partial and complete male sterility is often related to apomixis. In this study, we compared the morphology of pollen grains over several developmental stages in Miconia species with different rates of male sterility. Our aim was to improve the knowledge of mechanisms that lead to male sterility in this ecologically important tropical group. Routine techniques for microscopy were used to examine anthers in several developmental stages collected from the apomictic species Miconia albicans and M. stenostachya. Both species are completely male sterile since even the pollen grains with apparently normal cytoplasm were not able to develop a pollen tube. Meiosis is a rare event in M. albicans anthers and happens in an irregular way in M. stenostachya, leading to the pollen abortion. M. albicans has more severe abnormalities than M. stenostachya since even the microspores and pollen grain walls were affected. Moreover, in M. stenostachya, most mitosis occurring during microgametogenesis was also abnormal, leading to the formation of bicellular pollen grains with two similar cells, in addition to the formation of pollen grains of different sizes. Notably, abnormalities in both species did not reach the production of Übisch bodies, suggesting little or no tapetum involvement in male sterility in these two species.

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