A new angiosperms molecular specimen treatment method for field use

doi:10.17521/cjpe.2016.0322

Abstract

Abstract:

Aims This study aims to identify a more convenient drying method for obtaining molecular specimen of angiosperms in the field than the conventional silica gel drying method. Methods The leaves of Prunus serrulata var. lannesiana and Liriope spicata were dried under temperatures of 150 °C, 80 °C, 40 °C as well as under natural conditions, and by the silica gel drying method, respectively. The DNA extracts of various specimens were then analyzed using techniques of spectrophotometer detection, electrophoresis and PCR to evaluate the impacts of different drying treatments to the genomic DNA of testing plants. Important findings The concentrations of total DNA were higher for the specimens dried at 40 °C treatment and by the silica gel drying method than other treatments when assessed by the techniques of spectrophotometer detection and electrophoresis. The concentration of PCR products was highest in the specimens dried at 40 °C. Based on the results, the 40 °C drying can be recommended for obtaining molecular specimens of angiosperms because of its minimum degree of degradation, for convenience of operation and avoiding carrying large amounts of silica gel in field investigations.

Key words: angiosperms, field, molecular specimen, DNA, drying, silica gel

Feng-Jiao SHEN, Qian-Qian REN, Qi DONG, Li ZHU, Jian-Fang ZHANG, Jing YANG, Ran ZHANG, Hong-Zhu LIANG, Jian-Cheng ZHAO, Shuo SHI. A new angiosperms molecular specimen treatment method for field use[J]. Chin J Plan Ecolo, 2017, 41(7): 787-794.

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URL: https://www.plant-ecology.com/EN/10.17521/cjpe.2016.0322

https://www.plant-ecology.com/EN/Y2017/V41/I7/787

Figures/Tables 5

Table 1 Comparisons of DNA purity, concentration and the concentration of PCR products in the specimens of Prunus serrulata var. lannesiana and Liriope spicata obtained with different drying methods (mean ± SD)

干燥方式 Drying method	日本晚樱 Prunus serrulata var. lannesiana			山麦冬 Liriope spicata
干燥方式 Drying method	OD260/280	DNA (ng·μL^-1)	PCR (ng·μL^-1)	OD260/280	DNA (ng·μL^-1)	PCR (ng·μL^-1)
150	1.83 ± 0.17^a	451.24 ± 150.91^a	13.94 ± 3.97^c	2.03 ± 0.04^a	211.43 ± 64.31^a	0.74 ± 2.30^d
80	1.80 ± 0.13^a	376.13 ± 121.04^a	34.48 ± 9.42^b	1.83 ± 0.16^b	80.64 ± 48.16^c	16.51 ± 17.49^c
40	1.74 ± 0.16^ab	470.44 ± 228.50^a	49.07 ± 4.83^a	1.90 ± 0.08^ab	241.41 ± 88.07^a	57.64 ± 6.51^a
Y	1.73 ± 0.09^ab	291.64 ± 90.77^b	38.38 ± 4.75^b	1.93 ± 0.14^ab	160.01 ± 22.95^b	46.98 ± 8.22^b
S	1.64 ± 0.10^b	432.59 ± 167.67^a	39.95 ± 5.82^b	1.90 ± 0.24^ab	252.33 ± 61.74^a	43.19 ± 8.72^b

Fig. 1 Genomic DNA in specimens of Prunus serrulata var. lannesiana (R) obtained with five different drying methods. 150, drying at 150 °C; 80, drying at 80 °C; 40, drying at 40 °C; Y, natural drying; S, silica gel drying; 2K, 2 kb plus DNA ladder.

Fig. 2 Genomic DNA in specimens of Liriope spicata (M) obtained with five different drying methods. 50, drying at 150 °C; 80, drying at 80 °C; 40, drying at 40 °C; Y, natural drying; S, silica gel drying; 2K, 2 kb plus DNA ladder.

Fig. 3 Agarose gel electrophoresis of PCR products in specimens of Prunus serrulata var. lannesiana (R) obtained with five different drying methods. 150, drying at 150 °C; 80, drying at 80 °C; 40, drying at 40 °C; Y, natural drying; S, silica gel drying; 2K, 2kb plus DNA ladder.

Fig. 4 Agarose gel electrophoresis of PCR products in specimens of Liriope spicata (M) obtained with five different drying methods. 150, drying at 150 °C; 80, drying at 80 °C; 40, drying at 40 °C; Y, natural drying; S, silica gel drying; 2K, 2 kb plus DNA ladder.

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