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Gossypiumbarbadenseis a cultivated cotton species and possesses many desirable traits, including high fiber quality and resistance to pathogens, especiallyVerticillium  dahliae(a devastating pathogen of   Gossypium hirsutum  , the main cultivated species). These elite traits are difficult to be introduced into   G. hirsutum   through classical breeding methods. In addition, genetic transformation ofG.  barbadensehas not been successfully performed. It is therefore important to develop methods for evaluating the function and molecular mechanism of genes inG.  barbadense    . In this study, we had successfully introduced a virus-induced gene silencing (VIGS) system into three cultivars ofG.  barbadenseby inserting marker genes into the tobacco rattle virus (TRV) vector. After we optimized the VIGS conditions, including light intensity, photoperiod, seedling age and Agrobacterium strain, 100% of plants agroinfiltrated with the  GaPDS  silencing vector showed white colored leaves. Three other marker genes,  GaCLA1 ,  GaANS  and  GaANR , were employed to further test this VIGS system inG.  barbadense    . The transcript levels of the endogenous genes in the silenced plants were reduced by more than 99% compared to control plants; these plants presented phenotypic symptoms 2 weeks after inoculation. We introduced a fusing sequence fragment of  GaPDS  and  GaANR  gene silencing vectors into a single plant, which resulted in both photobleaching and brownish coloration. The extent of silencing in plants agroinfiltrated with fusing two-gene-silencing vector was consistent with plants harboring a single gene silencing vector. The development of this VIGS system should promote analysis of gene function inG.  barbadense    , and help to contribute desirable traits for breeding ofG.  barbadenseand   G. hirsutum  .

Development of Agrobacterium-Mediated Virus-Induced Gene Silencing and Performance Evaluation of Four Marker Genes in Gossypium barbadense