Expression of TUBB4B in mouse primary spermatocyte GC-2 cells and its regulatory effect on NF-κB and MAPK signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the interaction between Tubulin beta 4B class IVb (TUBB4B) and Agtpbp1/cytosolic carboxypeptidase- like1 (CCP1) in mouse primary spermatocytes (GC-2 cells) and the role of TUBB4B in regulating the development of GC-2 cells.@*METHODS@#Lentiviral vectors were used to infect GC-2 cells to construct TUBB4B knockdown and negative control (NC-KD) cells. The stable cell lines with TUBB4B overexpression (Tubb4b-OE) and the negative control (NC-OE) cells were screened using purinomycin. RT-qPCR and Western blotting were used to verify successful cell modeling and explore the relationship between TUBB4B and CCP1 expressions in GC-2 cells. The effects of TUBB4B silencing and overexpression on the proliferation and cell cycle of GC-2 cells were evaluated using CCK8 assay and flow cytometry. The signaling pathway proteins showing significant changes in response to TUBB4B silencing or overexpression were identified using Western blotting and immunofluorescence assay and then labeled for verification at the cellular level.@*RESULTS@#Both TUBB4B silencing and overexpression in GC-2 cells caused consistent changes in the mRNA and protein expressions of CCP1 (P < 0.05). Similarly, TUBB4B expression also showed consistent changes at the mRNA and protein after CCP1 knockdown and restoration (P < 0.05). TUBB4B knockdown and overexpression had no significant effect on proliferation rate or cell cycle of GC-2 cells, but caused significant changes in the key proteins of the nuclear factor kappa-B (NF-κB) signaling pathway (p65 and p-p65) and the mitogen-activated protein kinase (MAPK) signaling pathway (ErK1/2 and p-Erk1/2) (P < 0.05); CCP1 knockdown induced significant changes in PolyE expression in GC-2 cells (P < 0.05).@*CONCLUSIONS@#TUBB4B and CCP1 interact via a mutual positive regulation mechanism in GC-2 cells. CCP-1 can deglutamize TUBB4B, and the latter is involved in the regulation of NF-κB and MAPK signaling pathways in primary spermatocytes.

Anther Transcriptomics and Expression Analysis of Genes Related to Anthocyanin Synthesis in Blue Labeled Genic Male Sterile Wheat / 中国生物化学与分子生物学报

In order to reveal the molecular mechanism of blue labeled genic male sterility (BM-type GMS) and utilize the heterosis of BM-type GMS, we used the anthers of white-seed plants WS (sterile) and light blue seed plants WF (normal fertility) as experimental materials to analyze the differences in gene expression between them by transcriptome technology. And we also verified the genes expressed in anthocyanin synthesis in this study. Compared with WF, a total of 2352 differentially expressed genes were detected in WS. According to GO functional annotation, these genes could be divided into 3 categories and 43 subgroups. They are mainly involved in biosynthesis, phenylpropane metabolism, L-phenylalanine catabolism, membrane components, plasma membrane, cytoplasm, ATP binding, protein serine/threonine kinase activity, etc. KEGG pathway analysis showed that there were 159 genes enriched in the phenylpropanoid biosynthesis pathway, followed by the phenylalanine pathway, including 136 differentially expressed genes. Other genes are also involved a variety of amino acid metabolism, purine metabolism, pyrimidine metabolism and sugar metabolism pathway. Related to anthocyanin metabolism, several structural genes of key enzymes were differentially expressed, and most of them were up-regulated in WF, while only Flavanone 3-hydroxylase (F3H) and colorless anthocyanin dioxygenase (ANS) were down-regulated. Quantitative real-time PCR showed that the expression of 10 genes related to anthocyanin metabolism had the same trend as that in transcriptome sequencing data. Sequence homology analysis showed that the two selected transcription factors (DN48762c2g1 and DN25944c0g1) are clustered into the same cluster as the transcription factors regulating anthocyanin biosynthesis in maize, rice and Arabidopsis thaliana, which might be candidate genes for the blue aleurone layer of light blue seed plants in wheat. And fluorescence quantitative analysis showed that the expression level of DN48762c2g1 and DN25944c0g1 in WF was significantly higher than that in WS. In conclusion, the genes related to the anthocyanin biosynthesis pathway are not only related to the blue grain trait, but also may be involved in the anther abortion of BM-type GMS.

Ubiquitinome Profiling Reveals the Landscape of Ubiquitination Regulation in Rice Young Panicles / 基因组蛋白质组与生物信息学报·英文版

Ubiquitination, an essential post-transcriptional modification (PTM), plays a vital role in nearly every biological process, including development and growth. Despite its functions in plant reproductive development, its targets in rice panicles remain unclear. In this study, we used proteome-wide profiling of lysine ubiquitination in rice (O. sativa ssp. indica) young panicles. We created the largest ubiquitinome dataset in rice to date, identifying 1638 lysine ubiquitination sites on 916 unique proteins. We detected three conserved ubiquitination motifs, noting that acidic glutamic acid (E) and aspartic acid (D) were most frequently present around ubiquitinated lysine. Enrichment analysis of Gene Ontology (GO) annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of these ubiquitinated proteins revealed that ubiquitination plays an important role in fundamental cellular processes in rice young panicles. Interestingly, enrichment analysis of protein domains indicated that ubiquitination was enriched on a variety of receptor-like kinases and cytoplasmic tyrosine and serine-threonine kinases. Furthermore, we analyzed the crosstalk between ubiquitination, acetylation, and succinylation, and constructed a potential protein interaction network within our rice ubiquitinome. Moreover, we identified ubiquitinated proteins related to pollen and grain development, indicating that ubiquitination may play a critical role in the physiological functions in young panicles. Taken together, we reported the most comprehensive lysine ubiquitinome in rice so far, and used it to reveal the functional role of lysine ubiquitination in rice young panicles.

Development of KASP marker for cytoplasmic male sterility in Nicotiana tabacum and utilization in trait introgression

Cytoplasmic male sterility (CMS) is an important trait for large-scale hybrid seed production which avoids manual emasculation and undesired horizontal spread of pollen.Rearrangements in mitochondrial genome in terms of deletions and insertions are frequent causes leading to CMS. Mitochondrial ATP synthase is a multisubunit molecular machine which is involved in synthesis of ATP. In this study, three mutations in ATPase subunit 6 were identified and their cosegregation with male sterility was established using tobacco male sterile hybrids and Nicotiana suvaolensis. A breeder friendly Kompetitive allele specific polymerase chain reaction (KASP) SNP marker was developed for high throughput and quick genotyping. Introgression of this trait into selected germplasm lines (n = 9) was achieved based on foreground for CMS and background selection for recurrent parent using KASP marker and 50K custom tobacco SNP array, respectively. Analysis of genotyping data from SNP array revealed the presence of 88–99% of recurrent parent genome in BC3F1 plants. The selected BC3F1 plants exhibit CMS and are indistinguishable from the fertile recurrent parent (germplasm) in terms of plant morphology.

Nucleotide sequence polymorphism in the RFL-PPR genes of potato

Cytoplasmic male sterility (CMS) is widely used for hybrid seed production in cultivated Solanaceae species. However, there is very limited information about CMS-Rf genetic systems in potato (Solanum tuberosum). Studying the CMS-Rf systems in potato is both of theoretical and practical significance due to the emergence of a new revolutionary strategy of reinventing potato as adiploid inbred line-based crop to develop F1 hybrid seed potato breeding (Lindhout et al. 2011; Jansky et al. 2016). To search for potato Rf gene candidates, the comparative genetic approach was applied. Based on similarity to petunia Rf-PPR592 gene, 38 fragments were identified in five loci of the whole-genome nucleotide sequence of the accession DM 1-3 516 R44 S. tuberosum Phureja group (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The putative encoded mitochondrial proteins have 589–597 amino acid residues, similarto RF-PPR proteins of petunia and chili pepper and contain 14 or 15 PPR motifs. Primers have been developed flanking the most variable 782–865 bp regions of the selected loci, and polymorphism of the cloned fragments has been investigated in a subset of nine potato genotypes. The amplified fragments included seven or eight PPR motifs and lacked introns. The SNP frequencies ranged from 7.0 to 19.8% depending on the locus, while the ratio of nonsynonymous to synonymous substitutions varied between 0.9 and 2.1.Positions 1, 3 and 6 were the most variable in the studied PPR motifs. Our results demonstrated that the analysed sequences belong to the RFL-PPR gene subfamily and may be considered as Rf gene candidates in potato.

RNA editing analysis of ATP synthase genes in the cotton cytoplasmic male sterile line H276A

BACKGROUND: Pollen development is an energy-consuming process that particularly occurs during meiosis. Low levels of adenosine triphosphate (ATP) may cause cell death, resulting in CMS (cytoplasmic male sterility). DNA sequence differences in ATP synthase genes have been revealed between the N- and S-cytoplasms in the cotton CMS system. However, very few data are available at the RNA level. In this study, we compared five ATP synthase genes in the H276A, H276B and fertile F1 (H276A/H268) lines using RNA editing, RNA blotting and quantitative real time-PCR (qRT-PCR) to explore their contribution to CMS. A molecular marker for identifying male sterile cytoplasm (MSC) was also developed. RESULTS: RNA blotting revealed the absence of any novel orf for the ATP synthase gene sequence in the three lines. Forty-one RNA editing sites were identified in the coding sequences. RNA editing showed that proteins had 32.43% higher hydrophobicity and that 39.02% of RNA editing sites had proline converted to leucine. Two new stop codons were detected in atp6 and atp9 by RNA editing. Real-time qRT-PCR data showed that the atp1, atp6, atp8, and atp9 genes had substantially lower expression levels in H276A compared with those in H276B. By contrast, the expression levels of all five genes were increased in F1 (H276A/H268). Moreover, a molecular marker based on a 6-bp deletion upstream of atp8 in H276A was developed to identify male sterile cytoplasm (MSC) in cotton. CONCLUSIONS: Our data substantially contributes to the understanding of the function of ATP synthase genes in cotton CMS. Therefore, we suggest that ATP synthase genes might be an indirect cause of cotton CMS. Further research is needed to investigate the relationship among ATP synthase genes in cotton CMS.

Screening of different AFLP fragments between near-isogenic lines of male sterile and fertile Salvia miltiorrhiza and their comparison analysis / 中国中药杂志

There is distinctive advantage of using male sterile lines to breed new cultivar and produce hybrids, when compared with general breeding method on yield and quality. In our previous work, near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained through continuous hybridization in many years. In this investigation, 378 primer combination were screened by using AFLP and BSA technique, in which 26 markers amplified from seven primers were found to tightly link to male sterile gene. Based on these markers, two linkage genetic maps were constructed. A 2 027,2 028 bp fragment was amplifed from NILs of fertile and sterile S. miltiorrhiza, respectively, using genome walking technique and previous E11/M4-208 marker as template. Four base mutations were found in intron when comparing both fragments. Among all different markers between NILs of male sterile and fertile S. miltiorrhiza, four was found to have 100% identities to chromosome 1, 3 and 5 of Arabidopsis, namely, E01/M09-418, E05/M13-308, E05/M04-750 and E01/M01-204. The E01/M09-418 marker was very close to male sterile gene of S. miltiorrhiza with distance of 2.1 cM, which also had 100% identities to male sterile gene MS2 in Arabidopsis. Both were distributed in chromosome 3 of Arabidopsis. The 2 028 bp fragment also had 100% identities to MS2 gene. Another E05/M04-750 marker that had 100% identities to chromosome 5 of Arabidopsis was found to have high identities to POP085-M05 gene of poplars and low affinity calcium antiporter CAX2 of Arabidopsis with very low E-value. The constructed genetic map and differential fragments with potential functions found in this study provide a solid foundation to lock male sterile genes in S. miltiorrhiza genome and to discover their functions.

The value of scrotal ultrasonography in diagnosis of male sterility / 中国基层医药

Objective To investigate the value of scrotal ultrasonography in diagnosis of male sterility.Methods The scrotal ultrasonographic features of 162 cases of male sterility were retrospectively analyzed.Results The diagnostic accuracy of scrotal ultrasonography was 69.14％ (112/162) ; scrotal ultrasonography abnormalities were detected in 64 cases of varicocele (34.7％),showed tortuous ducts in spermatic cord and regurgitation in sonogmphy ;testicular microlithiasis disease in 16 cases,13 cases of testicular microlithiasis,36 cases of epididymal cysts,6 cases of hydrocele testis,testicular cyst 5 cases,7 cases of spermduct became thickened.Conclusion Scrotal ultrasonography can clearly show the structure of scrotum and has great value in the diagnosis of male infertility,while ultrasound has a cheap,convenient,high reproducibility and no damage,no pain.It is worthy of wide promotion and application.

The change of regular semen parameters in infertile patients with simple low-pH semen before and after symptomatic treatment / 国际检验医学杂志

Objective To study the change of regular semen parameters in infertile patients with simple low-pH semen before and after symptomatic treatment.Methods A total of 100 male infertility patients were recruited in the study,and were divided into two groups according to the pH range:patients group Ⅰ(n=40)and patients group Ⅱ(n=60);The two patients groups were tea-ted for low pH semen.When the first and second course of treatment were finished,semen pH values and conventional semen pa-rameters were tested and compared with those before treatment and control group.Results When the first course of treatment were finished,semen pH value of 75%(30/40)patients in group Ⅰ reached normal range,compared with the control group,the semen parameters were statistically different(P 0.05);Semen pH value of 90%(54/60)patients in group Ⅱ reached normal range,compared with the control group,the semen parameters and pH values were not statistically different(P >0.05 ). Conclusion The semen quality improved after the symptomatic treatment for low semen pH value and its an effective therapeutic approach for infertile patients with simple low-pH semen.

Analysis of pulse features in 57 patients of oligoathenoteratospermia / 国际中医中药杂志

ObjectiveTo investigate the TCM pathogenesis of male infertility through analyzing the characteristics of the pulse of oligoathenoteratospermia patients.MethodsRetrospect analysis was performed to get the pulse characteristics of oligoathenoteratospermia patients.The relation between the regularity of pulse and oligoathenoteratospermia was studied.ResultsThe pulse of oligoathenoteratospermia patients mainly presented at the both guan pulses and the both chi pulses,especially at the left guan pulse and the right chi pulse with thready pulse,wiry pulse,slippery pulse,uneven pulse,and moderate pulse,besides there were some differences of pulse between oligospermia and asthenozoospermia.ConclusionThe liver and kidney were the main location of oligoathenoteratospermia,and the main pathogenesis was asthenia of kidney,blood stasis,and dampness-heat.

Desbalance redox en la infertilidad masculina / Lack of Redox balance in male sterility

La infertilidad masculina es considerada como uno de los factores que más contribuyen a la infertilidad de la pareja. En conjunción a los factores convencionales que la originan se ha identificado una causa de extraordinaria repercusión: el estrés oxidativo, el cual es el resultado del desequilibrio entre la generación de especies reactivas del oxígeno y los antioxidantes, que puede originar daños o deformidades a los espermatozoides y eventualmente infertilidad masculina. Las especies reactivas del oxígeno se producen por diversos mecanismos en el semen, por espermatozoides inmóviles o con problemas de movilidad, leucocitos y por espermatozoides normales morfológicamente pero funcionalmente anormales. Entre estos daños, se registran la peroxidación lipídica a la membrana espermática y la fragmentación del ADN tanto nuclear como mitocondrial. Los daños que causa el estrés oxidativo sobre el ADN pueden originar trastornos en la descendencia que incluyen cáncer y acondroplastia. La peroxidación lipídica, origina la alteración de la membrana que afecta su estructura y función. La membrana del espermatozoide humano contiene una elevada proporción de ácidos grasos poliinsaturados, por lo tanto su susceptibilidad a la peroxidación lipídica es muy elevada. Las especies reactivas del oxígeno tienen diversos efectos sobre las células espermáticas, constituyendo un tema muy controvertido, por lo que este artículo tuvo como propósito actualizar al lector sobre algunos de los aspectos bioquímicos relacionados con la producción de especies reactivas del oxígeno y los métodos de diagnóstico que se emplean para detectarlo en la infertilidad humana.

The male sterility is considered as one of the factors more contributing to couple sterility. Join to conventional factors originate it condition it was possible to identify a cause with a high level of repercussion: the oxidative stress, which is the result of the imbalance between generation of reactive oxygen species and the antioxidants that may to cause damages or deformities in the spermatozoa and eventually male sterility. The reactive oxygen species are produced due to different mechanisms present in semen, motionless spermatozoa or with motility disorders, leukocytes and morphologically normal spermatozoa but functionally abnormal. Among these damages it is present the lipid peroxidation to spermatic membrane and the nuclear and mitochondrial DNA fragmentation. The damages of oxidative stress on DNA may also to cause disorders in offspring including cancer and achondroplasia. Lipid peroxidation leads to an alteration in the membrane affecting its structure and function. The human spermatozoon membrane contains a high proportion of poly-non-saturated fatty acids therefore its oversensitivity to lipid peroxidation is very high. The reactive oxygen species have different effects on spermatic cells, being a very controversial topic. The aim of present paper was to update readers on some of the biochemical features related to the production of reactive oxygen species and the diagnostic methods used to detect the human sterility.

Analysis of genetic diversity for wolfberry germplasms by AFLP technology / 中草药

Objective: To study the genetic relationships of fifteen wolfberry germplasm, including seven wild species, three varieties, and five cultivars and lines. Methods: Data were analyzed by the technology of genomic DNA-AFLP markers and the NTSYS average similarity cluster analysis. Results: The statistics showed that 432 bands were amplified using eight pairs of primers, polymorphic bands had account of 360 numbers, and polymorphic rate was 83.3%. All germplasm were classified into nine groups, five groups and three groups as different coefficient of 0.72,0.79, and 0.85. Conclusion: This study reveals a diverse basis and their genetic relationship of Chinese wolfberry at the molecular level. The male sterility YX-1 is clustered together with a white flowers wolfberry and a round fruit wolfberry, and YX-1 is more close to the two kinds of wolfberry but farther to the Ningxia wolfberry in the genetic distant. In addition, the classification is obviously different with the traditional morphological one about three variants.

Restoring pollen fertility in transgenic male-sterile eggplant by Cre/loxp-mediated site-specific recombination system

This study was designed to control plant fertility by cell lethal gene Barnase expressing at specific developmental stage and in specific tissue of male organ under the control of Cre/loxP system, for heterosis breeding, producing hybrid seed of eggplant. The Barnase-coding region was flanked by loxP recognition sites for Cre-recombinase. The eggplant inbred/pure line ('E-38') was transformed with Cre gene and the inbred/pure line ('E-8') was transformed with the Barnase gene situated between loxp. The experiments were done separately, by means of Agrobacterium co-culture. Four T0-plants with the Barnase gene were obtained, all proved to be male-sterile and incapable of producing viable pollen. Flowers stamens were shorter, but the vegetative phenotype was similar to wild-type. Five T0-plants with the Cre gene developed well, blossomed out and set fruit normally. The crossing of male-sterile Barnase-plants with Cre expression transgenic eggplants resulted in site-specific excision with the male-sterile plants producing normal fruits. With the Barnase was excised, pollen fertility was fully restored in the hybrids. The phenotype of these restored plants was the same as that of the wild-type. Thus, the Barnase and Cre genes were capable of stable inheritance and expression in progenies of transgenic plants.

Evidence of programmed cell death during microsporogenesis in an interspecific Brachiaria (Poaceae: Panicoideae: Paniceae) hybrid

Morphological changes have been investigated during plant programmed cell death (PCD) in the last few years due to the new interest in a possible apoptotic-like phenomenon existing in plants. Although PCD has been reported in several tissues and specialized cells in plants, there have been few reports of its occurrence during microsporogenesis. The present study reports a typical process of PCD during meiosis in an interspecific Brachiaria hybrid leading to male sterility. In this hybrid, some inflorescences initiated meiosis but it was arrested in zygotene/pachytene. From this stage, meiocytes underwent a severe alteration in shape showing substantial membrane blebbing; the cytoplasm became denser at the periphery; the cell nucleus entered a progressive stage of chromatin disintegration, and then the nucleolus disintegrated, and the cytoplasm condensed and shrunk. The oldest flowers of the raceme showed only the callose wall in the anthers showing obvious signs of complete sterility.

cDNA-AFLP Reveals Variation of Sequence Region trnF-ndhJ in Cytoplasmic Male Sterile Chinese Cabbage-pak-choi / 生物化学与生物物理进展

By means of cDNA amplified fragment length polymorphism(cDNA-AFLP) technique, a fragment P1708 was amplified from Polima cytoplasmic male sterile Chinese cabbage-pak-choi (Brassica campestris L. ssp. chinenesis Makino, syn, B. rapa L. ssp. chinenesis) 'Bpol97-05A'. RT-PCR showed that this fragment was specifically expressed in male sterile material. Sequencing and BLAST search in GenBank database indicated that P1708 had 100% homolog with chloroplast ndhJ-trnF gene region except a 54 bp insertion. Gene specific primer pairs were synthesized according to ndhJ-trnF gene region and two fragments about 1 900 bp were amplified respectively using genomic DNA templates of Polima cabbage and male fertile oilseed rape. The sequencing results showed that the gene region ndhJ-trnF of Polima cabbage contained two 54 bp repeats and some variation sites. The repeat part shared the same sequence as trnF gene except three bases at 5′ ends. For the insertion of 108 bp sequence, a new open reading frame was created.

An exploration of relationship between the infection of Ureaplasma Urealyticum and Chlamydia Trachomatis and male sterility / 中国艾滋病性病

Objective To discuss the relationship between the infection of Ureaplasma Urealyticum(Uu) and Chlamydia trachomatis(Ct) and male sterility.Methods To detect 476 specimens of sperm by FQ-PCR,326 specimens were collected from sterile men(sterility group) and 150 specimens from normal men(control group).Results The total detection rate of Uu and Ct was 49.69% in sterility group,being significantly higher than 16.67% in control group(?~2=46.98,P

Evaluation of the courtship and of the hybrid male sterility among Drosophila buzzatii cluster species (Diptera, Drosophilidae)

In the Drosophila repleta group the establishment of subgroups and complexes made on the basis of morphological and cytological evidences is supported by tests of reproductive isolation. Among species in the repleta group, the buzzatii cluster, due to its polymorphism and polytipism, is an excellent material for ecological and speciation studies. Some interspecific crosses involving Drosophila seriema, Drosophila sp. B, D. koepferae and D. buzzatii strains were completely sterile while others involving strains from these species produced F1 hybrids that did not yield F2. In the present work, data on courtship duration and copula occurrence obtained in the analysis of flies from parental sterile crosses and on spermatozoon mobility observed in F1 hybrids that did not yield F2 are presented. Copula did not occur during one hour of observation and the spermatozoon also did not show mobility at any of the analyzed stages (3, 7, 9 and 10 days old). There was a high variation in courtship average duration and in the percentage of males that courted the females. The reproductive isolation mechanisms indicated by these observations were pre and post-zygotic, as supported by the absence of copula and male sterility. Data obtained also showed the occurrence of different degrees of reproductive compatibility among the strains classified as the same species but from distinct geographic localities.

No grupo repleta de Drosophila, o estabelecimento de subgrupos e complexos realizado com base em evidências citológicas e morfológicas é suportado por testes de isolamento reprodutivo. Entre as espécies do grupo repleta, o cluster buzzatii, devido a seu politipismo e polimorfismo, é um excelente material para estudos ecológicos e de especiação. Alguns cruzamentos interespecíficos envolvendo linhagens de Drosophila seriema, Drosophila sp. B, D. koepferae e D. buzzatii foram completamente estéreis, enquanto outros produziram híbridos F1 que não deixaram F2. No presente trabalho são apresentados dados sobre a duração da corte e ocorrência de cópula dos intercruzamentos estéreis e dados sobre a mobilidade dos espermatozóides dos híbridos F1 que não deixaram F2. Não houve ocorrência de cópula no período de 1 hora de observação e os espermatozóides dos híbridos analisados não foram móveis em qualquer das idades testadas (3, 7, 9 e 10 dias de idade). Houve alta variação na duração média da corte e na porcentagem dos machos que cortejaram as fêmeas. A ausência de cópula e esterilidade dos machos F1 indica mecanismos pré e pós-zigótico operando entre as espécies desse cluster. Os dados também mostram diferentes níveis de compatibilidade reprodutiva entre linhagens classificadas como da mesma espécie mas de diferentes localidades geográficas.

Agronomic performance of male-sterile and fertile maize genotypes at two plant populations / Performace agronômica de genótipos macho-estéreis e férteis de milho em duas densidades de semeadura

This experiment was conducted in 1994 at Ames, Iowa, US to test whether cytoplasmic male-sterility can be used to decrease barrenness and to increase grain yield of maize at two plant populations. Four genotypes were tested: a hybrid (NK 6330) and an inbred, wifh sterile and fertile counterparts. Each genotype owas evaluated at plant populations equivalent to 25,000 and 75,000 pl. ha-1. Hybrids produced higher grain yield than inbreds at both plant populations. Gram yield was higher at 75,000 than at 25,000 pl. ha-1. No difference in gram yield, number of ears per plant, number of grains per ear, tassel length, and tassel number of branches was found between sterile and fertile counterparts of the inbred and hybrid, regardiess of plant population. Fertile genotypes bore heavier tassels at anthesis than their sterile counterparts. Adequate precipitation distribution and high fertility level in the soil probably decreased competition between tassel and ears, mitigating potential yield benefits of suppressing genetically pollen production.

Este experimento foi conduzido em Ames, Iowa, US durante o ano agrícola de 1994, tendo como objetivo avaliar se a macho-esterelidade genético-citoplasmática pode ser utilizada para aumentar o rendimento de grãos de milho em diferentes populações de planta. Quatro genótipos foram utilizados: um híbrido (NK 6330) e uma linhagem, ambos em suas versões fértil e macho-estéril. Cada genótipo foi avaliado em duas densidades de semeadura, equivalentes a 25.000 e 75.000 pl.ha-1. Os híbridos produziram maiores rendimentos de grão do que as linhagens nas duas populações utilizadas. O rendimento de grãos por área foi maior em 75.000 do que em 25.000 pl.ha-1. Nenhuma diferença significativa em termos de rendimento de grãos, número de espigas por planta, número de grãos por espiga, comprimento e número de ramos do pendão, foi observada entre genótipos férteis e macho-estéreis, independentemente da população de plantas. As versões férteis apresentaram pendões mais pesados do que os genótipos macho-estéreis. A adequada distribuição da precipitação e a alta fertilidade do solo possivelmente diminuíram a competição entre o pendão e as espigas, minimizando os benefícios da supressão genética da produção de pólen sobre o rendimento de grãos.

Mitochondrial ATP synthase genes may be implicated in cytoplasmic male sterility in Sorghum bicolor.

Incompatible nuclear-cytoplasmic interactions are responsible for the phenomenon of cytoplasmic male sterility in plants. We have analysed male sterile (2077A, 296A), maintainer fertile (2077B, 296B) and fertility restored (2077R, 296R) lines of sorghum for the restriction fragment locations of various mitochondrial genes and their transcripts. We report here a polymorphism in genes related to the ATP synthase complex between two different cytoplasms from the A and Β set of lines of 2077 and 296. There is also a difference in the transcript size of the atpA gene between the A and Β cytoplasms. We propose that incompatibility in nuclear cytoplasmic interactions may be explained in terms of incompatible subunits being synthesized by the mitochondria and nucleus for a multisubunit complex of the mitochondrial membrane such as ATPase.

Mitochondrial genome organization and cytoplasmic male sterility in plants.

Plant mitochondrial genomes are much larger and more complex than those of other eukaryotic organisms. They contain a very active recombination system and have a multipartite genome organization with a master circle resolving into two or more subgenomic circles by recombination through repeated sequences. Their protein coding capacity is very low and is comparable to that of animal and fungal systems. Several subunits of mitochondrial functional complexes, a complete set of tRNAs and 26S, 18S and 5S rRNAs are coded by the plant mitochondrial genome. The protein coding genes contain group II introns. The organelle genome contains stretches of DNA sequences homologous to chloroplast DNA. It also contains actively transcribed DNA sequences having open reading frames. Plasmid like DNA molecules are found in mitochondria of some plants Cytoplasmic male sterility in plants, characterized by failure to produce functional pollen grains, is a maternally inherited trait. This phenomenon has been found in many species of plants and is conveniently used for hybrid plant production. The genetic determinants for cytoplasmic male sterility reside in the mitochondrial genome. Some species of plants exhibit more than one type of cytoplasmic male sterility. Several nuclear genes are known to control expression of cytoplasmic male sterility. Different cytoplasmic male sterility types are distinguished by their specific nuclear genes (rfs) which restore pollen fertility. Cytoplasmic male sterility types are also characterized by mitochondrial DNA restriction fragment length polymorphism patterns, variations in mitochondrial RNAs, differences in protein synthetic profiles, differences in sensitivity to fungal toxins and insecticides, presence of plasmid DNAs or RNAs and also presence of certain unique sequences in the genome. Recently nuclear male sterility systems based on (i) over expression of agrobacterial rol C gene and (ii) anther specific expression of an RNase gene have been developed in tobacco and Brassica by genetic engineering methods.