Creating a novel petal regeneration system for function identification of colour gene of grape hyacinth

Background Grape hyacinth (Muscari spp.) is one of the most important ornamental bulbous plants. However, its lengthy juvenile period and time-consuming transformation approaches under the available protocols impedes the functional characterisation of its genes in flower tissues. In vitro flower organogenesis has long been used to hasten the breeding cycle of plants but has not been exploited for shortening the period of gene transformation and characterisation in flowers. Results A petal regeneration system was established for stable transformation and function identification of colour gene in grape hyacinth. By culturing on Murashige and Skoog medium (MS) with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 8.88 μM 6-benzyladenine (6-BA), during the colour-changing period, the flower bud explants gave rise to regeneration petals in less than 3 months, instead of the 3 years required in field-grown plants. By combining this system with Agrobacterium-mediated transformation, a glucuronidase reporter gene (GUS) was delivered into grape hyacinth petals. Ultimately, 214 transgenic petals were regenerated from 24 resistant explants. PCR and GUS quantitative analyses confirmed that these putative transgenic petals have stably overexpressed GUS genes. Furthermore, an RNAi vector of the anthocyanidin 3-O-glucosyltransferase gene (MaGT) was integrated into grape hyacinth petals using the same strategy. Compared with the non-transgenic controls, reduced expression of the MaGT occurred in all transgenic petals, which caused pigmentation loss by repressing anthocyanin accumulation. Conclusion The Agrobacterium transformation method via petal organogenesis of grape hyacinth took only 3–4 months to implement, and was faster and easier to perform than other gene-overexpressing or -silencing techniques that are currently available.

Effect of PMEL17 Plumage Colour Gene Diversity on Production Performance of Indigenous Chicken Variety of Bangladesh

Background: An adaptive meat and egg type indigenous chicken is crucial for countries those depends on rural poultry production for meeting the protein requirements of the peoples. Genetic characterizations of native chickens have been documented, however, no study has observed the plumage colouration and its potential role in production traits. Thus, the aim of the current study was to know the effect of PME17, plumage colour gene diversity on production performance of indigenous chicken varieties.Methods: The plumage colours, comb and body shape of chickens corresponds with the live weight and egg production (clutch size) and the egg characteristics were recorded. Gel electrophoresis and polymerase chain reactions (PCR) were performed from blood cell DNA following standard protocols. The PCR products were sequenced using Sanger sequencing and for molecular analysis MEGA6 software were used. Result: Highest live weight (1400±25.4 g) and egg production (15.3±0.9 /number /clutch) was obtained in spotted-single-round chicken than other varieties. Both external and internal egg characteristics differed between varieties and spotted- single-round variety found to be best than other varieties. The sequence of PMEL17 gene was 99% homology with the sequence of Gallus gallus and Gallus gallus domesticus. A mutation was observed at 91bp nucleotide in brownish and at 64bp positional nucleotide and in black-white chicken variety.

AFLP and SSR markers linked to the yellow seed colour gene in Brassica juncea L.

Yellow mustard, cultivated in northern Shaanxi of China, is a valuable germplasm of Brassica juncea with low erucic acid content. Its yellow seed colour is controlled by a recessive allele of a single gene, whose dominant allele conditions brown seed colour. To map the yellow seed colour allele, amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) technologies were used to identify markers linked to the recessive allele. The analysis was done on 386 F₂ plants, segregating for seed colour, from the cross Wuqi yellow mustard × Wugong mustard. The plants were selfed to determine their seed colour genotype. Twenty AFLP markers and eight SSR markers were identified from 256 AFLP primer combinations and 624 pairs of SSR primers, respectively. Blast analysis indicated that the sequences of four closely linked AFLP and SSR markers showed good collinearity with those of Arabidopsis chromosome 3, and the homologue of the yellow seed colour allele was located between At3g14190 and At3g32130. Sequence information of the region between the two genes of Arabidopsis could be used to develop more closely linked markers to narrow down the homologue of the yellow seed colour allele. These results would accelerate the procedure of yellow seed colour gene cloning and marker-assisted selection for yellow mustard.