Spatiotemporal gene expression profiles associated with male strobilus development in Cryptomeria japonica by suppression subtractive hybridization

Abstract

To identify the genes associated with the formation and development of the male strobilus of Cryptomeria japonica D. Don (Sugi), male strobilus-specific suppression subtractive hybridization (SSH) libraries were constructed from three different stages of male strobilus development. From these SSH libraries, 1,012 unigene sets including 314 contigs were assembled from 2,448 sequences. The profiles of the genes isolated from the SSH libraries differed strongly according to developmental stage: 49%, 13% and 29% of genes were unique to the early stage (stage E), tetrad stage (stage T), and mature stage (stage M) SSH libraries, respectively. To evaluate the reliability of the SSH libraries, we focused on the eight genes strongly concentrated in the three SSH libraries and performed RT-PCR analysis. All genes tested were expressed more strongly in the male strobilus than in the shoot. Our findings suggest that the gene profiles associated with male strobilus development differ in both number and kind of dominantly expressed genes.

A protocol for <i>Agrobacterium</i>-mediated transformation of Japanese cedar, Sugi (<i>Cryptomeria japonica</i> D. Don) using embryogenic tissue explants

“…Genetic transformation, and genome editing technology based on it, can be a powerful tool for shortening the breeding period of trees whose generations are many years apart. A large number of expressed sequence tags obtained from vegetative buds, inner bark, female strobili, male strobili, pollen, sapwood, xylem, cambium, and roots, have been identified (Fukuda et al 2018;Futamura et al 2006Futamura et al , 2008Kurita et al 2011;Mishima et al 2014Mishima et al , 2018Nose and Watanabe 2014;Tsumura et al 1997;Ueno et al 2019;Ujino-Ihara et al 2000Yoshida et al 2007Yoshida et al , 2012, and various genetic linkage maps and DNA markers have been developed (summarized by Tsumura 2011). Genetic transformation is also an essential tool for taking advantage of these abundant resources for molecular breeding and identification of gene function by reverse genetic analysis.…”

Inheritance of male flower production in Cryptomeria japonica (sugi) estimated from analysis of a diallel mating test

“…In Japan, approximately 20 % of adults suffer from C. japonica and Chamaecyparis obtusa pollinosis (Okuda 2003), which has become a serious health problem. Many studies have been performed on the flowering of C. japonica, such as histological analysis (Hashizume 1962), estimation of clonal variation of flower production (Saito and Kawasaki 1984), and recently, genetic analysis including establishment of cDNA or expressed sequence tag libraries (Futamura et al 2008;Kurita et al 2011) from male reproductive organs.…”

High-efficiency Agrobacterium-mediated transformation of Cryptomeria japonica D. Don by co-cultivation on filter paper wicks followed by meropenem treatment to eliminate Agrobacterium

“…Furthermore, there have been recent reports on ESTs from male strobilus-specific suppression subtractive hybridization libraries and the xylem transition zone (Kurita et al 2011;Yoshida et al 2012). Genetic transformation is also an essential tool that facilitates the effective use of these extensive resources for reverse genetic studies and molecular breeding.…”