The tomato (Solanum lycopersicum) male sterile 32 (ms32) mutant has been used in hybrid seed breeding programs largely because it produces no pollen and has exserted stigmas. In this study, histological examination of anthers revealed dysfunctional pollen and tapetum development in the ms32 mutant. The ms32 locus was fine mapped to a 28.5 kb interval that encoded four putative genes. Solyc01g081100, a homolog of Arabidopsis bHLH10/89/90 and rice EAT1, was proposed to be the candidate gene of MS32 because it contained a single nucleotide polymorphism (SNP) that led to the formation of a premature stop codon. A codominant derived cleaved amplified polymorphic sequence (dCAPS) marker, MS32D, was developed based on the SNP. Real-time quantitative reverse-transcription PCR showed that most of the genes, which were proposed to be involved in pollen and tapetum development in tomato, were downregulated in the ms32 mutant. These findings may aid in marker-assisted selection of ms32 in hybrid breeding programs and facilitate studies on the regulatory mechanisms of pollen and tapetum development in tomato.

Broccoli (Brassica oleracea var. italica) is an important commercial vegetable crop. As part of an efficient pollination system, cytoplasmic male sterility (CMS) has been widely used for broccoli hybrid production. Identifying the original sources of CMS in broccoli accessions has become an important part of broccoli breeding. In this study, the diversity of the CMS sources of 39 broccoli accessions, including 19 CMS lines and 20 hybrids, were analyzed using mitochondrial markers. All CMS accessions contained the ogu orf138-related DNA fragment and the key genes of nap CMS, pol CMS, and tour CMS were not detected. The 39 CMS accessions were divided into five groups using six orf138-related and two simple sequence repeat markers. We observed that ogu CMS R3 constituted 79.49% of the CMS sources. CMS6 and CMS26 were differentiated from the other accessions using a specific primer. CMS32 was distinguished from the other accessions based on a 78-nucleotide deletion at the same locus as the orf138-related sequence. When the coefficient was about 0.90, five CMS accessions (13CMS6, 13CMS23, 13CMS24, 13CMS37, and 13CMS39) exhibiting abnormal floral organs with poor seed setting were grouped together. The polymerase chain reaction amplification profiles for these five accessions differed from those of the other accessions. We identified eight useful molecular markers that can be used to detect CMS types during broccoli breeding. Our data also provide important information relevant to future studies on the possible origins and molecular mechanisms of CMS in broccoli.

Bud abortion is the main factor affecting hybrid seeds’ yield during broccoli cross breeding when using ogura cytoplasmic male sterile (ogu CMS) lines. However, the genes associated with bud abortion are poorly understood. We applied RNA sequencing to analyze the transcriptomes of normal and abortive buds of broccoli maintainer and ogu CMS lines. Functional analysis showed that among the 54,753 annotated unigenes obtained, 74 and 21 differentially expressed genes in common were upregulated and downregulated in ogu CMS abortive buds compared with ogu CMS normal buds, maintainer normal, and abortive buds, respectively. Nineteen of the common differentially expressed genes were enriched by GO terms associated with glycosyl hydrolases, reactive oxygen species scavenging, inhibitor, and protein degradation. Ethylene-responsive transcription factor 115 and transcriptional factor basic helix-loop-helix 137 were significantly upregulated; transcription factors DUO1 and PosF21/RF2a/BZIP34 were downregulated in ogu CMS abortive buds compared with the other groups. Genes related to polygalacturonase metabolism, glycosyl hydrolases, oxidation reduction process, phenylalanine metabolism, and phenylpropanoid biosynthesis were significantly changed in ogu CMS abortive buds. Our results increase our understanding of bud abortion, provide a valuable resource for further functional characterization of ogu CMS during bud abortion, and will aid in future cross breeding of Brassica crops.