Dataset Information

Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla.

ABSTRACT: The northern root-knot nematode (Meloidogyne hapla) is a critical pathogen with a wide host range. Quantitative real-time polymerase chain reaction (qRT-PCR) has been used to elucidate gene expression and function of M. hapla. Suitable reference genes are required to ensure accurate results of qRT-PCR for normalising gene expression. Eleven candidate reference genes of M. hapla were selected to evaluate gene expression stability under different conditions. The stability of candidate reference genes was ranked using RefFinder analysis, and the optimal number of reference genes was recommended with geNorm. Notably, the most stable reference genes were SDHA, Mdh, and RpS6 for all samples; SDHA and RpS6 were particularly stable during development stage treatments, whereas Mdh and RpS6 were appropriate for temperature and inorganic compound treatments. In contrast, the least stable reference genes were Actin1 during development stages and all other treatments, GAPDH for temperature treatments, and ?-Tub for inorganic compound treatments. One target gene, Mh-Hsp90, was used to verify the selection of reference genes, results showed Mdh and RpS6 could be used as suitable reference genes for M. hapla, and Mdh plus RpS6 were better. Our finding contributes to further work on gene transcription analysis in M. hapla.

SUBMITTER: Wu X

PROVIDER: S-EPMC6583968 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla.

Wu Xiaojing X Yu Hongyan H Yang Rouwei R Zhou Yuanyuan Y Zhu Xiaofeng X Wang Yuanyuan Y Liu Xiaoyu X Fan Haiyan H Chen Lijie L Duan Yuxi Y

PloS one 20190619 6

The northern root-knot nematode (Meloidogyne hapla) is a critical pathogen with a wide host range. Quantitative real-time polymerase chain reaction (qRT-PCR) has been used to elucidate gene expression and function of M. hapla. Suitable reference genes are required to ensure accurate results of qRT-PCR for normalising gene expression. Eleven candidate reference genes of M. hapla were selected to evaluate gene expression stability under different conditions. The stability of candidate reference ge ...[more]

PMID: 31216347

Similar Datasets

Project description:Root-knot nematodes (RKN) are the most serious plant parasitic nematodes having a broad host range exceeding 2,000 plant species. Quercus brantii Lindl. and Q. infectoria Oliv are the most important woody species of Zagros forests in west of Iran where favors sub-Mediterranean climate. National Botanical Garden of Iran (NBGI) is scheduled to be the basic center for research and education of botany in Iran. This garden, located in west of Tehran, was established in 1968 with an area of about 150 ha at altitude of 1,320 m. The Zagros collection has about 3-ha area and it has been designed for showing a small pattern of natural Zagros forests in west of Iran. Brant's oak (Q. brantii) and oak manna tree (Q. infectoria) are the main woody species in Zagros collection, which have been planted in 1989. A nematological survey on Zagros forest collection in NBGI revealed heavily infection of 24-yr-old Q. brantii and Q. infectoria to RKN, Meloidogyne hapla. The roots contained prominent galls along with egg sac on the surface of each gall. The galls were relatively small and in some parts of root several galls were conjugated, and all galls contained large transparent egg masses. The identification of M. hapla was confirmed by morphological and morphometric characters and amplification of D2-D3 expansion segments of 28S rRNA gene. The obtained sequences of large-subunit rRNA gene from M. hapla was submitted to the GenBank database under the accession number KP319025. The sequence was compared with those of M. hapla deposited in GenBank using the BLAST homology search program and showed 99% similarity with those KJ755183, GQ130139, DQ328685, and KJ645428. The second stage juveniles of M. hapla isolated from Brant's oak (Q. Brantii) showed the following morphometric characters: (n = 12), L = 394 ± 39.3 (348 to 450) µm; a = 30.9 ± 4 (24.4 to 37.6); b = 4.6 ± 0.44 (4 to 5.1); b΄ = 3.3 ± 0.3 (2.7 to 3.7), c = 8.0 ± 1 (6.2 to 10.3), ć = 5.3 ± 0.8 (3.5 to 6.3); Stylet = 12.1 ± 0.8 (11 to 13) µm; Tail = 50 ± 5.6 (42 to 57) µm; Hyaline 15 ± 1.8 (12 to 18) µm. Oak manna, Q. infectoria population of second stage juveniles clearly possessed short body length and consequently other morphometric features were less than those determined for Q. brantii population, and these features were: (n = 12), L = 359.0 ± 17.3 (319 to 372) µm; a = 28.6 ± 3 (22.8 to 31); b = 5.0 ± 0.3 (4.8 to 5.2); b΄ = 3.3 ± 0.2 (3 to 3.6), c = 8.1 ± 0.5 (7.4 to 8.8), ć = 4.7 ± 0.5 (3.9 to 5.2); Stylet = 11.4 ± 0.7 (10 to 12) µm; Tail = 44 ± 1.8 (42 to 47) µm; Hyaline 12 ± 1.7 (10 to 15) µm. To date two species of Meloidogyne, M. quercianaGolden, 1979 and M. christieiGolden and Kaplan, 1986 have been reported to parasitize oaks (Quercus spp.) from the United States of America. M. querciana was found on pin oak Quercus palustris in Virginia. The oak RKN infected pine oak, red oak, and American chestnut heavily in greenhouse tests (Golden, 1979). The other species M. christiei was described from turkey oak and Q. laevis in Florida, which has monospecific host range (Golden and Kaplan, 1986). Both of these RKN species seem to be restricted to the United States of America and have not been reported from other place. According to our knowledge this is the first report of occurrence of M. hapla on Q. brantii and Q. infectoria in the world. This study includes these two oak species to the host range of RKN, M. hapla for the world and expands the information of RKN, M. hapla host ranges on oaks.

Dataset Information

Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla.

Publications

Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets