Identification and characterization of fungal pathogens associated with black root rot on saffron from Southern Khorasan, Iran

Javad Ramezani Avval Reiabi — 2024-02-09

Saffron, a spice derived from the dried stigmas of Crocus sativus flowers, is renowned for its high cost and culinary value. As a perennial herbaceous plant belonging to the Iridaceae family, saffron is susceptible to diseases such as corm rot, which has previously been attributed to various fungal pathogens including Fusarium, Penicillium, and Botrytis. It has been observed that soil-borne fungi are consumed by the bulb mite. Nonetheless, a significant amount of saprophytic fungi can be found in the soil of saffron fields. This may lead one to wonder if the saffron corm mite is a main or secondary pest. In this study, we aimed to identify the underlying causes of the early yellowing of saffron leaves and the reduction in the quantity and quality of this valuable product. A total of 100 plant samples displaying symptoms of infection, along with their corms, were collected and analyzed from various saffron cultivation areas in Qaen City, Southern Khorasan Province. Our findings revealed that the fungal genera Rhizoctonia and Curvularia were the primary causes of saffron root rot. This is the first report of Curvularia associated with saffron in Iran and other countries. The benefits of the disease-related information presented here extend to a diverse group of stakeholders, including farmers, learners, researchers, plant protection organizations, development departments, extension workers, policymakers, government agencies, and public organizations.

Construction of Mambalgin-1 gene cassette for transient expression in apoplastic space

Ghaffar Khezri Khezri — 2024-04-18

Mambalgine-1 is a protein that acts by blocking ASICs channels in neurons as a potent analgesic such as morphine while having no adverse effects of morphine, such as addiction and respiratory distress. In this study, in order to produce inexpensive and scalable Mambalgin-1 protein, recombinant PVX-Mambalgin-1 viral vector was designed and optimized for transient expression in apoplastic space of Nicotiana benthamiana. To compensate for the lack of methionine at the beginning of Mambalgin-1 and also secretion of this protein into apopalstic space Glucanase inhibitor protein 2 (GIP2) signal peptide was added to the N-terminal of Mambalgin-1. The 6xhis-tag was added to the C-terminal of Mambalgin-1 for downstream processes and protein purification. Probability of signal peptide cleavage and secretory of Mambalgin-1 assessed by SignalP.5 server. The desired protein was reverse translated into nucleotide sequences based on N. benthamiana Codon Usage from Kazusa Database; then mRNA destabilizing sequences, polyadenylation signal and hidden stop codons were modified using Visual Gene Developer 1.9 and Mega4 softwares. CAI and GC were kept as high as possible. ClaI/SalI restriction enzyme sites were added at the 3’ and 5’ end of the construct. The results showed that the GIP2 signal peptide would most likely (0.96) be cleaved from Mambalgin-1 and Mambalgin-1 expression will be highly secretory (0.998) into the apopalstic space after expression. After optimization, cloning and transfection of Agrobacterium, the existence and transformation of recombinant PVX-Mambalgin-1 vector was confirmed by colony PCR.

Journal of Applied Plant Biology

Identification and characterization of fungal pathogens associated with black root rot on saffron from Southern Khorasan, Iran

Construction of Mambalgin-1 gene cassette for transient expression in apoplastic space