Improvement of cotton fiber quality by transforming the acsA and acsB genes into Gossypium hirsutum L. by means of vacuum infiltration

Abstract

A novel method for the genetic transformation of cotton pollen by means of vacuum infiltration and Agrobacterium-mediated transformation is reported. The acsA and acsB genes, which are involved in cellulose synthesis in Acetobacter xylinum, were transferred into pollen grains of brown cotton with the aim of improving its fiber quality by incorporating useful prokaryotic features into the colored cotton plants. Transformation was carried out in cotton pollen-germinating medium, and transformation was mediated by vector pCAMBIA1301, which contains a reporter gene beta-glucuronidase (GUS), a selectable marker gene, hpt, for hygromycin resistance and the genes of interest, acsA and acsB. The integration and expression of acsA, acsB and GUS in the genome of transgenic plants were analyzed with Southern blot hybridization, PCR, histochemical GUS assay and Northern blot hybridization. We found that following pollination on the cotton stigma transformed pollen retained its capability of double-fertilization and that normal cotton seeds were produced in the cotton ovary. Of 1,039 seeds from 312 bolls pollinated with transformed pollen grains, 17 were able to germinate and grow into seedlings for more than 3 weeks in a nutrient medium containing 50 mg/l hygromycin; eight of these were transgenic plants integrated with acsA and acsB, yielding a 0.77% transformation rate. Fiber strength and length from the most positive transformants was 15% greater than those of the control (non-transformed), a significant difference, as was cellulose content between the transformed and control plants. Our study suggests that transformation through vacuum infiltration and Agrobacterium mediated transformation can be an efficient way to introduce foreign genes into the cotton pollen grain and that cotton fiber quality can be improved with the incorporation of the prokaryotic genes acsA and acsB.

Dissecting Functions of KATANIN and WRINKLED1 in Cotton Fiber Development by Virus-Induced Gene Silencing

“…Our results are consistent with the data of Zhang et al (2011) that the increased fiber yield is associated with the cost of a smaller seed size, resulting from the repartition of seed oil into carbohydrate fibers. This result is consistent with previous reports that transgenic overexpression of carbohydrate biosynthesis genes improved cotton fiber length and strength (Li et al, 2004;Haigler et al, 2007;Jiang et al, 2012). All these studies suggest the possibility to increase fiber length by diverting the flow of fixed carbon from seed oil into carbohydrate fibers.…”

“…Since cellulose is the major constituent of cotton fiber, it is not surprising that several carbohydrate biosynthetic genes have been shown to modulate fiber development. Transgenic overexpression of a SUCROSE SYNTHASE gene, a SUCROSE PHOSPHATE SYNTHASE gene, and cellulose synthesis genes improved cotton fiber length and strength (Li et al, 2004;Haigler et al, 2007;Jiang et al, 2012). Similarly, higher xyloglucan endotransglycosylase/ hydrolase (XTH) activity can promote fiber cell elongation, and transgenic cotton overexpressing the xth gene has been shown to increase mature fiber length (Lee et al, 2010).…”

Molecular characterization and transcriptome profiling of expansin genes isolated from Calotropis procera fibers

“…The efforts to accomplish these goals have been carried out utilizing both the classical breeding and genetic engineering tools (Guo et al 2003;Ruan et al 2003;Li et al 2004;Arioli, 2005;Ruan et al 2005;Wilkins et al 2005;Momtaz and Soliman, 2008). The over expression of G. hirsutum expansin gene has been reported for a fourfold increase in fiber yield that was cumulatively due to the increase in fiber length, number and fiber weight per seed.…”

Isolation and characterization of gene sequences expressed in cotton fiber

“…The GhACT1 gene plays an essential role in various cellular processes, including cell division, elongation, differentiation and morphogenesis (WANG;CHEN;XIA, 2005). Another important account of modification of fiber characteristics using transgenics is quoted by Li et al (2004), who introduced the acsA and acsB genes involved in cellulose synthesis in Acetobacter xylinum, and obtained cotton plants whose fiber displayed superior characteristics for length, quality and cellulose content.…”