CMU Intellectual Repository: Genotypic characterization of arbuscular mycorrhizal fungi and their potential use for growth of lettuces = ลักษณะทางพันธุกรรมของเชื้อราอาร์บัสคูลาร์ไมตอร์ไรซาและศักยภาพต่อการเจริญเติบโตของผักกาดหอม / Maliwan Saouy

Thesis Title Genotypic Characterization of Arbuscular Mycorrhizal Fungi and Their Potential Use for Growth of Lettuces

Author Miss Maliwan Saouy

Degree Doctor of Philosophy (Biotechnology)

Thesis Advisory Committee Assoc. Prof. Dr. Somporn Choonluchanon Chairperson Dr. Arawan Shutsrirung Member Dr. Choochat Santasup Member

ABSTRACT

This study was aimed to collect the indigenous AM spores from lettuce cultivated soil for identification and selection for effective root infection. The selected spores were used for inoculum production and determination of lettuce yield and nutrients uptake. The preliminary investigation of AM fungal spores from lettuce rhizosphere was extracted by wet-sieving and decanting method. A total of 40 soil samples were collected from different vegetable farms. AM fungi were found only in the practice field of Department of Horticulture site at the average densities of 4.68 spores per 100 g dry soil. The characterization of AM fungi was identified in both morphological and molecular biology aspects. Molecular identification methods based on genomic DNA extraction from single spore and PCR analysis of the 18S rRNA genes were used for sequencing. The collected isolates were belong to Glomus sp.

(85% homology; AJ852533.1) and Gigasporasp. (70% homology; AB197869.1). The percentage of root colonization was evaluated in Lactuca sativar var crispa L.

(green leaf lettuce and red leaf lettuce) which were inoculated with Glomus sp. and Gigasporasp. After inoculation period of four weeks with 50 spores in sand substrate, the total percentage of root colonization was significantly greater on red leaf lettuce than green leaf lettuce and the Glomus sp. colonized in lettuce root was higher than

Gigasporasp. These spores were then propagated by pot culture with four generations of multiplication. It was found that the spore of Gigaspora sp. was not observed in the third generation. Whereas the average spores propagation of Glomus sp. were 24 spores per g dry sand.

The development of AM fungal inoculum by hydroponic culture with two nutrient media, modified Long Ashton (LANS) and modified Hoagland medium (HNS) were compared for culturing Glomus sp.on red leaf lettuce. After a pre-culture period of four weeks with 50 spores in sand substrate, the initial infection of mycorrhizal plants irrigated with either LANS or HNS were not significantly different. At four weeks after transplanting, the total percentage of root colonization was significantly greater in HNS medium than in LANS medium. The four nutrient media (solution-I, solution-II, solution-III and solution-IV) of modified Hoagland were used and replaced with fresh solution every week in a deep water culture technique to compare for root colonization and spore production. Four weeks after transplanting, plants in solution-I medium had higher percentage of mycorrhizal colonization and spore population than with other nutrients media. The spore production was significantly positive correlated with root colonization while it was negative correlation with EC, shoot and root biomass. The combined effects of fertilization levels (0%, 25%, 50%, 75% and 100% of recommended fertilizer) and AM inoculation on Lactuca sativar var crispaL. (green leaf, red leaf and red grana), Lactuca sativarvar capitataL. (head lettuce and asmerunda) andLactuca sativar var longifolia L.(cos) was determined for plant yield, root colonization, macro and micronutrient assimilation. The higher fertilization reduced root colonization but the highest fertilization (100%) absolutely inhibited AM infection of all tested plants. The dual application of AM spores with low fertilizer rates (0%, 25%, 50%) suggested to increase yield and the uptake of N, P, Ca,Mg and Mn.

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