Molecular Identification and Antifungal Properties of Four Thaumatin-like Proteins in Spruce (Picea likiangensis)

Abstract

Thaumatin-like proteins (TLPs) are involved in the plant defense response against pathogens, and most of them exhibit antifungal activity. However, the role of TLPs in pathogen-induced defense responses in spruce is not fully understood. In this study, four TLP genes encoding thaumatin-like protein, designated as PlTLP1–4, were isolated and identified from Picea likiangensis needles. Sequence analysis showed that PlTLP1, PlTLP3, and PlTLP4 contained 16 conserved cysteine residues, while PlTLP2 had only 10 conserved cysteine residues. qPCR analysis showed that PlTLPs were expressed in all tissues tested, PlTLP1, PlTLP3, and PlTLP4 had the highest expression levels in young fruits, while PlTLP2 had the highest expression levels in roots. In addition, the expression levels of four PlTLPs were significantly upregulated during infection by Lophodermium piceae. Four recombinant PlTLPs expressed in Escherichia coli exhibited obvious β-1,3-glucanase activity. The antifungal activity assay showed that four recombinant PlTLPs had significant inhibitory effects on the mycelial growth of L. piceae, Fusarium proliferatum, Botrytis cinerea, and Roussoella doimaesalongensis. Microscopic observation revealed that the recombinant PlTLP1–4 induced the morphological changes of the mycelia of L. piceae, and the recombinant PlTLP2 and PlTLP3 induced the morphological changes of the mycelia of F. proliferatum and R. doimaesalongensis, while all the recombinant PlTLPs had no obvious negative effect on the morphology of B. cinerea mycelium. These results suggest that PlTLP genes may play an important role in the defense response of P. likiangensis against L. piceae invasion.

Cloning and Characterization of Two Novel PR4 Genes from Picea asperata

“…Two primer pairs, PaPR4-a-Fq/PaPR4-a-Rq and PaPR4-b-Fq/PaPR4-b-Rq ( Supplementary Table S1 ), were designed for RT-qPCR based on the sequences of PaPR4-a and PaPR4-b . EF (GenBank ID: AJ132534.1) and TIF (GenBank ID: DR448953.1) were used as the internal reference genes ( Supplementary Table S1 ) [ 75 , 76 , 77 ]. Hieff UNICON ® Universal Blue qPCR SYBR Green Master Mix (Yeasen Biotechnology Co., Ltd., Shanghai, China) was used for RT-qPCR, and the experiments were performed in triplicate.…”

Applying molecular and genetic methods to trees and their fungal communities

“…A TLP gene was identified in a GWAS as one hub gene of importance for pine tree breeding (Ding et al 2022b). The antifungal activity of Picea TLPs is based on glucanase activity on fungal hyphal cell walls (Liu et al 2021(Liu et al , 2022.…”

Tree Genetics: Molecular and Functional Characterization of Genes

“…As a positive consequence of the new technical approaches in -omics research [1] and the continuous decrease in sequencing and other molecular biology techniques costs, the papers do not merely focus on model species. Indeed, six papers focus on broadleaved trees and four on conifers, including economically relevant species for a plethora of different uses, but also rare/endangered species [3] and ecologically important trees [4,5]. The variety of different species studied by the research papers belonging to this Special Issue perfectly respond to the invitation reported in [2], to extend the research fields in forest genetics to a broader number of species.…”

“…In particular, [6] analyzes Camellia oleifera, which is used for oil production; [7] Liriodendron sino-americanum, a relevant ornamental and afforestation species; and [3] Handeliodendron bodinieri, a rare and endangered species, which is also important for bio-energy, landscaping, and water and soil conservation. The of [4] focuses on Picea likiangensis, one of the dominant tree species in the subalpine forest southwest China, with an important ecological in soil and water and carbon storage; [8] reports findings on Pinus massoniana, a relevant timber well as loblolly and slash pines studied in [9]. Chosenia arbutifolia [10] is used as a raw material for technological aims and for landscape planting in China.…”