[논문]Identification of Candidate Transcripts Related to Drought Stress using Secondary Traits and qRT-PCR in Tropical Maize (Zea mays L.)

Kim, Hyo Chul; Song, Kitae; Moon, Jun-Cheol; Kim, Jae Yoon; Kim, Kyung-Hee; Lee, Byung-Moo

doi:10.7740/kjcs.2019.64.4.432

Identification of Candidate Transcripts Related to Drought Stress using Secondary Traits and qRT-PCR in Tropical Maize (Zea mays L.) 원문보기

Korean journal of crop science = 韓國作物學會誌, v.64 no.4, 2019년, pp.432 - 440

Kim, Hyo Chul (Department of Life Science, Dongguk University-Seoul) , Song, Kitae (Department of Life Science, Dongguk University-Seoul) , Moon, Jun-Cheol (Agriculture and Life Sciences Research Institute, Kangwon National University) , Kim, Jae Yoon (Department of Plant Resources, College of Industrial Science, Kongju National University) , Kim, Kyung-Hee (Department of Life Science, Dongguk University-Seoul) , Lee, Byung-Moo (Department of Life Science, Dongguk University-Seoul)

Abstract ▼ AI-Helper

Global climate change exerts adverse effects on maize production. Among abiotic stresses, drought stress during the tasseling stage (VT) can increase anthesis-silking intervals (ASI) and decrease yield. We performed an evaluation of ASI and yield using a drought-sensitive line (Ki3) and a drought-tolerant line (Ki11) to analyze the correlation with ASI and yield. Moreover, the de novo data of Ki11 were analyzed to find putative novel transcripts related todrought stress in tropical maize. A total of 182 transcripts, with a log2 ratio >1.5, were found by comparing drought conditions to a control. The top 40 transcripts of high expression levels in the de novo analysis were selected and analyzed with PCR. Of the 40 transcripts, six novel transcripts were detected by quantitative real-time PCR (qRT-PCR) using seedling and VT stage samples. Five transcripts (transcripts_1, 12, 34, 35, and 40) were up-regulated in the Ki11 shoot at seedling stage, and transcripts_1, 12, and 40 were up-regulated at the re-watering stage after 12 h of drought stress. The transcripts_32 and 34 were up-regulated at the VT stage. Hence, transcript_34 possibly plays a significant role in drought tolerance during the seedling and VT stages. The transcript_32 was identified as chloramphenicol acetyltransferase (CAT) by Pfam domain analysis. The function of the other transcripts remained unknown. Further characterization of these novel transcripts in genetic regulation will be of great value for the improvement of maize production.

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표/그림 (5)

표 Table 1. Primer sets of transcript and reference genes used for qRT-PCR.
그림 Fig. 1. Relative expression levels of Ki11 (tolerant) and Ki3 (sensitive) under control (C) and drought stress (D).
표 Table 2. Day to pollen (days), day to silking (days), and ASI (days) of different maize genotypes under control and drought conditions in 2015, 2016, and 2017.
그림 Fig. 2. Relative expression levels of Ki11 (resistant) and Ki3 (sensitive) under control (C), drought stress (6 h and 12 h), and re-watering (after 6 h and 12 h) in seedling stage.
표 Table 3. Number of grains per ear (n), 100-grain weight (g), and grain yield (g/plant) of different maize geno-types under control and drought conditions in 2017.

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제안 방법

In this study, we evaluated the secondary traits using drought sensitive line (Ki3) and drought tolerant line (Ki11). The de novo data of RNA-seq in Ki11 were analyzed by qRT-PCR to find novel transcripts related to the drought stress in tropical maize.
Hence, de novo sequence is consisted with not exist in reference genome. In this study, we obtained with novel six putative transcripts for drought stress. In previous studies, de novo assembly was conducted in wheat (Duan et al.
Re-watering group was recovered by water supply during 24 h after 6 h and 12 h drought stress. The shoot, root, and leaf samples were conducted with three biological replicates for quantitative real-time PCR (qRTPCR). The root samples only used 12 h drought stress.

대상 데이터

We conducted with analysis of transcripts by qRT-PCR using seedling and VT stage samples for the three biological replicates. Eight transcripts were analyzed by qRT-PCR, 6 transcripts (transcript_1, 12, 32, 34, 35, and40) were expressed in all samples. However, 2 transcripts (transcript_38 and39) did not express in the most of samples.
The plants were grown in a plastic pot 240 mm wide, 380 mm length and 140 mm high (about 13 L) filled with 10 L vermiculite. We supplied 1 L water at every days, the temperature and humidity were maintained at 25°C and 40% during 16 hours with light/8 hours with dark in a growth room.

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