Cloning, characterization and promoter analysis of S-RNase gene promoter from Chinese pear (Pyrus pyrifolia)

Liu, Xueying; Wuyun, Tana; Zeng, Hongyan

doi:10.1016/j.gene.2012.06.017

Cited by 5 publications

(1 citation statement)

References 25 publications

(36 reference statements)

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“…Numerous studies on the extension of the storage period of yakli pears have been performed, which indicated that the effects of the external environment at harvest as well as during storage and processing affect the aging of yali pears (2,3). A series of preservation measures were developed; however, these measures were not able to fundamentally control the aging of pears (4). Therefore, in order to overcome the challenge of extending the shelf-life of yali pears, it is required to generate novel varieties of yali pear by modifying their genes associated with storage-associated aging characteristics.…”

Section: Introductionmentioning

confidence: 99%

Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation

Dong²,

Zhang

2012

Molecular Medicine Reports

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The aim of the present study was to genetically modify plantlets of the Chinese yali pear to reduce their expression of ripening-associated 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) and therefore increase the shelf-life of the fruit. Primers were designed with selectivity for the conserved regions of published ACO gene sequences, and yali complementary DNA (cDNA) cloning was performed by reverse transcription quantitative polymerase chain reaction (PCR). The obtained cDNA fragment contained 831 base pairs, encoding 276 amino acid residues, and shared no less than 94% nucleotide sequence identity with other published ACO genes. The cDNA fragment was inversely inserted into a pBI121 expression vector, between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator, in order to construct the anti‑sense expression vector of the ACO gene; it was transfected into cultured yali plants using Agrobacterium LBA4404. Four independent transgenic lines of pear plantlets were obtained and validated by PCR analysis. A Southern blot assay revealed that there were three transgenic lines containing a single copy of exogenous gene and one line with double copies. The present study provided germplasm resources for the cultivation of novel storage varieties of pears, therefore providing a reference for further applications of anti‑sense RNA technology in the genetic improvement of pears and other fruit.

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Section: Introductionmentioning

confidence: 99%

Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation

Dong²,

Zhang

2012

Molecular Medicine Reports

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Characterization of 25 full-length S-RNase alleles, including flanking regions, from a pool of resequenced apple cultivars

et al. 2018

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Data obtained from Illumina resequencing of 63 apple cultivars were used to obtain full-length S-RNase sequences using a strategy based on both alignment and de novo assembly of reads. The reproductive biology of apple is regulated by the S-RNase-based gametophytic self-incompatibility system, that is genetically controlled by the single, multi-genic and multi-allelic S locus. Resequencing of apple cultivars provided a huge amount of genetic data, that can be aligned to the reference genome in order to characterize variation to a genome-wide level. However, this approach is not immediately adaptable to the S-locus, due to some peculiar features such as the high degree of polymorphism, lack of colinearity between haplotypes and extensive presence of repetitive elements. In this study we describe a dedicated procedure aimed at characterizing S-RNase alleles from resequenced cultivars. The S-genotype of 63 apple accessions is reported; the full length coding sequence was determined for the 25 S-RNase alleles present in the 63 resequenced cultivars; these included 10 previously incomplete sequences (S , S , S , S , S , S , S , S , S and S ). Moreover, sequence divergence clearly suggests that alleles S and S , proposed to be neutral variants of the same alleles, should be instead considered different specificities. The promoter sequences have also been analyzed, highlighting regions of homology conserved among all the alleles.

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Learning from nature: new approaches to the metabolic engineering of plant defense pathways

Jirschitzka

Mattern

Gershenzon

et al. 2013

Current Opinion in Biotechnology

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Cloning, characterization and promoter analysis of S-RNase gene promoter from Chinese pear (Pyrus pyrifolia)

Cited by 5 publications

References 25 publications

Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation

Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation

Characterization of 25 full-length S-RNase alleles, including flanking regions, from a pool of resequenced apple cultivars

Learning from nature: new approaches to the metabolic engineering of plant defense pathways

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