Abstract
Paulownia witches' broom (PaWB) caused by phytoplasma might result in devastating damage to the growth and wood production of Paulownia. To study the effect of phytoplasma on DNA sequence and to discover the genes related to PaWB occurrence, DNA polymorphisms and DNA methylation levels and patterns in PaWB seedlings, the ones treated with various concentration of methyl methane sulfonate (MMS) and healthy seedlings were investigated with amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MSAP). Our results indicated that PaWB seedlings recovered a normal morphology, similar to healthy seedlings, after treatment with more than 20 mg·L−1 MMS; Phytoplasma infection did not change the Paulownia genomic DNA sequence at AFLP level, but changed the global DNA methylation levels and patterns; Genes related to PaWB were discovered through MSAP and validated using quantitative real-time PCR (qRT-PCR). These results implied that changes of DNA methylation levels and patterns were closely related to the morphological changes of seedlings infected with phytoplasmas.
Highlights
Paulownia, one of the fastest growing trees in the world, is native to China
Amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MSAP) are efficient and reliable methods to detect DNA polymorphism and DNA methylation, respectively [20,21]. With these two approaches, the variations of DNA polymorphisms and DNA methylation in PaWB seedlings (PS), the ones treated with methyl methane sulfonate (MMS) and healthy seedlings (HS) were investigated in order to reveal the genes associated with Paulownia witches’ broom (PaWB)
Sequence of Paulownia seedlings, Paulownia DNA polymorphisms of PS, the ones treated with MMS and HS were detected with 96 pairs of amplified fragment length polymorphism (AFLP) primer combinations (Figure S1)
Summary
One of the fastest growing trees in the world, is native to China. In recent years, it has been introduced in many other countries [1]. Since the 1970s, abundant of researches about the way and route of pathogen transmission [6], the prevention and control of insect vectors [7,8], and physiological and biochemical variation of Paulownia during the occurrence of PaWB were carried out, and several metabolic pathways related to PaWB were reported by Liu et al [9] and Mou et al [10]. These researches are helpful to understand the interaction of Paulownia and phytoplasma, the molecular mechanism of PaWB occurrence is still remain elusive
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