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Minc03328 effector gene downregulation triggered by in planta RNAi strategy strongly reduced plant susceptibility to Meloidogyne incognita and suggests that Minc03328 gene is a promising target for the development of genetically engineered crops to improve plant tolerance to M. incognita. Meloidogyne incognita is the most economically important species of root-knot nematodes (RKN) and causes severe damage to crops worldwide. M. incognita secretes several effector proteins to suppress the host plant defense response, and manipulate the plant cell cycle and other plant processes facilitating its parasitism. Different secreted effector proteins have already been identified in M. incognita, but not all have been characterized or have had the confirmation of their involvement in nematode parasitism in their host plants. Herein, we characterized the Minc03328 (Minc3s00020g01299) effector gene, confirmed its higher expression in the early stages of M. incognita parasitism in plants, as well as the accumulation of the Minc03328 effector protein in subventral glands and its secretion. We also discuss the potential for simultaneous downregulation of its paralogue Minc3s00083g03984 gene. Using the in planta RNA interference strategy, Arabidopsis thaliana plants overexpressing double-stranded RNA (dsRNA) were generated to specifically targeting and downregulating the Minc03328 gene during nematode parasitism. Transgenic Minc03328-dsRNA lines that significantly downregulated Minc03328 gene expression during M. incognita parasitism were significantly less susceptible. The number of galls, egg masses, and [galls/egg masses] ratio were reduced in these transgenic lines by up to 85%, 90%, and 87%, respectively. Transgenic Minc03328-dsRNA lines showed the presence of fewer and smaller galls, indicating that parasitism was hindered. Overall, data herein strongly suggest that Minc03328 effector protein is important for M. incognita parasitism establishment. As well, the in planta Minc03328-dsRNA strategy demonstrated high biotechnological potential for developing crop species that could efficiently control RKN in the field. Made available in DSpace on 2022-10-03T14:05:21Z (GMT). No. of bitstreams: 1 Minc03328-effector-gene-downregulation-2022.pdf: 6010297 bytes, checksum: 16efd63e19d2622a2a4e30c8810f27cd (MD5) Previous issue date: 2022

Made available in DSpace on 2019-04-12T00:44:59Z (GMT). No. of bitstreams: 1 document.pdf: 1990174 bytes, checksum: d8eab807c485ac8924ec4d84f23f772c (MD5) Previous issue date: 2019 Na publicação: Maria Cristina Mattar Silva, Maria Fatima Grossi-de-Sa.

Gossypium hirsutum (commercial cooton) is one of the most economically important fibers sources and a commodity crop highly affected by insect pests and pathogens. Several transgenic approaches have been developed to improve cotton resistance to insect pests, through the transgenic expression of different factors, including Cry toxins, proteinase inhibitors, and toxic peptides, among others. In the present study, we developed transgenic cotton plants by fertilized floral buds injection (through the pollen-tube pathway technique) using an DNA expression cassette harboring the cry1Ia12 gene, driven by CaMV35S promoter.

In plants, WRKY proteins are a group of transcription factors existing as a gene superfamily that play important roles in regulation of defense response pathways. To assess the diversity of this protein family in coffee (Coffea spp.), data mining methods were used on a set of around 200,000 coffee expressed sequence tags. A total of 53 different putative WRKY genes were obtained, but only 22 unigenes encoding a protein with a WRKY domain were identified, eight of which are supported by full-length cDNA sequences. Alignment of WRKY domain sequences of the coffee unigenes and 72 Arabidopsis thaliana WRKY genes showed that the 22 coffee WRKY members were distributed among the main A. thaliana WRKY subgroups and shared conserved peptide domains. To assess the involvement of WRKY genes in coffee defense response pathways, their expression was analyzed under biotic (nematode and rust fungus infection), hormonal (salicylic acid, methyl-jasmonate), and wounding treatments, leaf senescence, and fruit development. Five members of WRKY groups IId and III were regulated only by pathogens and hormone treatments. Although a significant correlation of WRKY genes expression after MeJA and rust treatments was observed, expression of coffee genes involved in JA biosynthesis and lipoxygenase (EC 1.13.11.12) activity assays did not support the involvement of JA in the early coffee resistance responses to the rust pathogen. The five WRKY transcription factor members identified might play important roles as regulators of pathogen resistance responses and could be useful for improving coffee tolerance to various biotic stresses.

Made available in DSpace on 2018-05-30T00:47:28Z (GMT). No. of bitstreams: 1 ID239761.pdf: 3068337 bytes, checksum: 61e00e1c44c69e0940c584bd71ad1e5e (MD5) Previous issue date: 2004-03-08

Made available in DSpace on 2019-10-08T00:33:41Z (GMT). No. of bitstreams: 1 ID226061.pdf: 360358 bytes, checksum: f3976ffb11bae317fa30085e9e16b8ca (MD5) Previous issue date: 2001