Insertion and Deletion (InDel) are common features in genomes and are associated with genetic variation. The whole-genome re-sequencing data from two parents (X1 and X2) of the elite cucumber (Cucumis sativus) hybrid variety Lvmei No.1 was used for genome-wide InDel polymorphisms analysis. Obtained sequence reads were mapped to the genome reference sequence of Chinese fresh market type inbred line ‘9930’ and gaps conforming to InDel were pinpointed. Further, the level of cross-parents polymorphism among five pairs of cucumber breeding parents and their corresponding hybrid varieties were used for evaluating hybrid seeds purity test efficiency of InDel markers. A panel of 48 cucumber breeding lines was utilized for PCR amplification versatility and phylogenetic analysis of these markers. In total, 10,470 candidate InDel markers were identified for X1 and X2. Among these, 385 markers with more than 30 nucleotide difference were arbitrary chosen. These markers were selected for experimental resolvability through electrophoresis on an Agarose gel. Two hundred and eleven (211) accounting for 54.81% of markers could be validated as single and clear polymorphic pattern while 174 (45.19%) showed unclear or monomorphic genetic bands between X1 and X2. Cross-parents polymorphism evaluation recorded 68 (32.23%) of these markers, which were designated as cross-parents transferable (CPT) InDel markers. Interestingly, the marker InDel114 presented experimental transferability between cucumber and melon. A panel of 48 cucumber breeding lines including parents of Lvmei No. 1 subjected to PCR amplification versatility using CPT InDel markers successfully clustered them into fruit and common cucumber varieties based on phylogenetic analysis. It is worth noting that 16 of these markers were predominately associated to enzymatic activities in cucumber. These agarose-based InDel markers could constitute a valuable resource for hybrid seeds purity testing, germplasm classification and marker-assisted breeding in cucumber.
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SUMMARYDevelopment of partial inter-specific hybrid (PIH) rice is a promising approach for exploiting inter-specific heterosis between Oryza glaberrima and Oryza sativa. In the present study, the relationship between genetic distance (GD) of parental lines and yield performance of the PIHs was assessed using partial diallel crosses between three indica male sterile lines and 14 introgression lines (ILs) with different fragments of O. glaberrima genes. Twenty two out of the 42 PIHs expressed positive heterobeltiosis (i.e., the Fl hybrid showed superiority over the better parent in the target trait) for panicle number, spikelet number, thousand grain weight and grain yield/plant. The proportion of O. glaberrima genome in the ILs ranged from 0·03 to 0·41, as revealed by 16 informative simple sequence repeat markers. Significant positive correlations were found between the proportion of O. glaberrima genome of the ILs and the GD between the ILs and the three different female parents. Heterosis of spikelet number per panicle in the hybrids was positively proportional with the O. glaberrima genome content of the parental ILs, while that of fertile grain percentage was negatively proportional to the O. glaberrima genome proportion. On average, the PIHs with higher grain yield and highest heterobeltiosis were obtained from the ILs carrying between 0·15 and 0·30 of O. glaberrima genome. The results indicated that a small proportion (<0·15) of the O. glaberrima genome in the ILs might limit heterosis expression of spikelet number per panicle, while a very large proportion (>0·30) of the O. glaberrima genome decreased the grain filling percentage in the PIHs, thus an intermediate range of O. glaberrima genome proportion should be more suitable for breeding heterotic PIHs. The exploitation of inter-specific heterosis between O. glaberrima and O. sativa has potential value for heterotic breeding in rice.
The rice dwarf and narrow leaf mutant 2 (dnl2) is dwarfed and forms narrow and brittle leaves. Its dwarfness was shown to be due to its shortened internodes, resulting from a reduced size of the internode parenchyma cells. Its narrow and brittle leaves were attributed to a compromised ability to form vascular bundles but the reduced fiber content and a thin cortical layer. However, the response to the application of either gibberellin or brassinolide was not different between dnl2 and its wild type. Transcription profiling indicated that a number of cell division/expansion-associated and crude fiber synthesis-related genes were down-regulated in the mutant. A genetic analysis revealed that the mutant phenotype is under monogenic control, and the gene responsible was mapped to a 50.1 kb genomic region on the long arm of chromosome 10. This region was shown to harbour ten open reading frames. Although transcription profiling of these genes indicated that three were differentially transcribed in the mutant, there was no sequence polymorphism in the coding sequence between the mutant and the wild type alleles.Additional key words: fine-mapping, leaf and stem structure, mechanical strength, Oryza sativa, transcriptional analysis.
ABSTRACT. The dwarf and narrow-leaf rice (Oryza sativa L.) mutant dnl3 was isolated from the Japonica cultivar Zhonghua 11 (wildtype). dnl3 exhibited pleiotropic developmental defects. The narrowleaf phenotype resulted from a marked reduction in the number of vascular bundles, while the dwarf stature was caused by the formation of foreshortened internodes and a reduced number of parenchyma cells. The suggestion that cell division is impaired in the mutant was consistent with the transcriptional behavior of various genes associated with cell division. The mutant was less responsive to exogenously supplied gibberellic acid than the wild-type, and profiling the transcription of genes involved in gibberellin synthesis and response revealed that a Resequencing the open reading frames revealed that the mutant carried an allele at one of the three genes that differed from the wild-type sequence by 2-bp deletions; this gene encoded a cellulose synthase-like D4 (CSLD4) protein. Therefore, OsCSLD4 is a candidate gene for DNL3. DNL3 was expressed in all of the rice organs tested at the heading stage, particularly in the leaves, roots, and culms. These results suggest that DNL3 plays important roles in rice leaf morphogenesis and vegetative development.
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