Expression of Amaranthin-like Lectins Gene and Responses to Abiotic Stresses in Cucumber

doi:10.11983/CBB20177

Abstract

Abstract: Lectins are globular proteins with carbohydrate-binding sites which enable them to specifically recognize and bind single or more particular carbohydrate structures. Previous studies have shown that lectins play critical roles in plant defense against their herbivores and pathogens. Due to their toxicity towards organisms like bacteria, fungi, viruses and insects, lectins are believed to have great potential in both agricultural and medical applications. As one of the smallest lectin families, there has been very limited research on amaranthin-like lectins. Here, we analyzed the distributions and intron/exon structures from 16 different amaranthin-like genes in the genome of cucumber, an important economic crop. To evaluate their responses against different stresses/plant hormones, promoter analysis was performed for all amaranthin-like genes. Finally, real-time quantitative PCR were performed on stress-treated plants to analyze the responses of amaranthin-like genes towards cold, salt, drought stresses and ABA treatment. This work provides valuable information for the study of physiological roles of amaranthin-like proteins and their involvement in plant defense.

Key words: abiotic stress, amaranthin-like lectins, promoter, protein domain

Fei Zhao, Liuyi Dang, Minhui Wei, Chunying Liu, Wei Leng, Chenjing Shang. Expression of Amaranthin-like Lectins Gene and Responses to Abiotic Stresses in Cucumber[J]. Chinese Bulletin of Botany, 2021, 56(2): 183-190.

Add to citation manager EndNote|Ris|BibTeX

URL: https://www.chinbullbotany.com/EN/10.11983/CBB20177

https://www.chinbullbotany.com/EN/Y2021/V56/I2/183

Figures/Tables 7

References 22

[1]	王梦龙, 彭小群, 陈竹锋, 唐晓艳 (2020). 植物凝集素类受体蛋白激酶研究进展. 植物学报 55,96-105.
[2]	王志斌, 张秀梅, 郭三堆 (2000). 在转基因植物中利用植物凝集素防治害虫的研究. 植物学通报 17,108-113.
[3]	曾日中, 黎瑜 (1998). 橡胶蛋白—一种与胶乳凝固有关的具有抗真菌活性的植物凝集素. 植物学通报 15(增刊),24-28.
[4]	Al Atalah B, Fouquaert E, Van Damme EJM (2013). Promoter analysis for three types of EUL-related rice lectins in transgenic Arabidopsis. Plant Mol Biol Rep 31, 1315- 1324.
[5]	Chen SC, Jin WJ, Liu AR, Zhang SJ, Liu DL, Wang FH, Lin XM, He CX (2013). Arbuscular mycorrhizal fungi (AMF) increase growth and secondary metabolism in cucumber subjected to low temperature stress. Sci Hortic 160,222-229.
[6]	Dang LY, Rougé P, Van Damme EJM (2017). Amaranthin-like proteins with aerolysin domains in plants. Front Plant Sci 8,1368.
[7]	Dang LY, Van Damme EJM (2016). Genome-wide identification and domain organization of lectin domains in cucumber. Plant Physiol Biochem 108,165-176.
[8]	Faruque K, Begam R, Deyholos MK (2015). The amaranthin-like lectin (LuALL) genes of flax: a unique gene family with members inducible by defence hormones. Plant Mol Biol Rep 33,731-741.
[9]	Ghazarian H, Idoni B, Oppenheimer SB (2011). A glycobiology review: carbohydrates, lectins and implications in cancer therapeutics. Acta Histochem 113,236-247.
[10]	Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999). Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Res 27,297-300.
[11]	Huang SW, Li RQ, Zhang ZH, Li L, Gu XF, Fan W, Lucas WJ, Wang XW, Xie BY, Ni PX, Ren YY, Zhu HM, Li J, Lin K, Jin WW, Fei ZJ, Li GC, Staub J, Kilian A, van der Vossen EAG, Wu Y, Guo J, He J, Jia ZQ, Ren Y, Tian G, Lu Y, Ruan J, Qian WB, Wang MW, Huang QF, Li B, Xuan ZL, Cao JJ, Asan, Wu ZG, Zhang JB, Cai QL, Bai YQ, Zhao BW, Han YH, Li Y, Li XF, Wang SH, Shi QX, Liu SQ, Cho WK, Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng ZC, Zhang SP, Wu J, Yang YH, Kang HX, Li M, Liang HQ, Ren XL, Shi ZB, Wen M, Jian M, Yang HL, Zhang GJ, Yang ZT, Chen R, Liu SF, Li JW, Ma LJ, Liu H, Zhou Y, Zhao J, Fang XD, Li GQ, Fang L, Li YR, Liu DY, Zheng HK, Zhang Y, Qin N, Li Z, Yang GH, Yang S, Bolund L, Kristiansen K, Zheng HC, Li SC, Zhang XQ, Yang HM, Wang J, Sun RF, Zhang BX, Jiang SZ, Wang J, Du YC, Li SG (2009). The genome of the cucumber, Cucumis sativus L. Nat Genet 41, 1275- 1281.
[12]	Li YM, Li SH, He XR, Jiang WL, Zhang DL, Liu BB, Li QM (2020). CO₂ enrichment enhanced drought resistance by regulating growth, hydraulic conductivity and phytohormone contents in the root of cucumber seedlings. Plant Physiol Biochem 152,62-71.
[13]	Migocka M, Papierniak A (2011). Identification of suitable reference genes for studying gene expression in cucumber plants subjected to abiotic stress and growth regulators. Mol Breed 28,343-357.
[14]	Pfaffl MW, Horgan GW, Dempfle L (2002). Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30,e36.
[15]	Shang CJ, Dang LY, Van Damme EJM (2017). Plant AB toxins with lectin domains. In: Gopalakrishnakone P, Carlini RC, Ligabue-Braun R, eds. Plant Toxins. Dordrecht: Springer. pp.1-14.
[16]	Szczesny P, Iacovache I, Muszewska A, Ginalski K, van der Goot FG, Grynberg M (2011). Extending the aerolysin family: from bacteria to vertebrates. PLoS One 6,e20349.
[17]	Tsaneva M, Van Damme EJM (2020). 130 years of plant lectin research. Glycoconj J 37,533-551.
[18]	Tuteja N (2007). Abscisic acid and abiotic stress signaling. Plant Signal Behav 2,135-138.
[19]	Van Holle S, Van Damme EJM (2019). Messages from the past: new insights in plant lectin evolution. Front Plant Sci 10,36.
[20]	Wang J, Pan CT, Wang Y, Ye L, Wu J, Chen LF, Zou T, Lu G (2015). Genome-wide identification of MAPK, MAPKK, and MAPKKK gene families and transcriptional profiling analysis during development and stress response in cucumber. BMC Genomics 16,386.
[21]	Yan SS, Che G, Ding L, Chen ZJ, Liu XF, Wang HY, Zhao WS, Ning K, Zhao JY, Tesfamichael K, Wang Q, Zhang XL (2016). Different cucumber CsYUC genes regulate response to abiotic stresses and flower development. Sci Rep 6,20760.
[22]	Zhu YX, Jia JH, Yang L, Xia YC, Zhang HL, Jia JB, Zhou R, Nie PY, Yin JL, Ma DF, Liu LC (2019). Identification of cucumber circular RNAs responsive to salt stress. BMC Plant Biol 19,164.

Primer name	Forward primer (5'-3')	Reverse primer (5'-3')
CACS	TGGGAAGATTCTTA- TGAAGTGC	CTCGTCAAATTT- ACACATTGGT
PP2A	CAACAGGTGATATT- GGATTATGAT	GCCAGCTCATCC- TCATATAAG
AAT4	TTCGTACACGCAAC- GAGA	TGAAGAGGGTAA- GGCTTG
NAAT1	GAGGAGCTGTGAAA- GGAGCA	CCCTCCACGACA- GTTCCAAT
AAT9	CAGAAACAGCGAAC- CAGAGC	AACTTCATCCCCA- CCGAGTT
AAT14	GGGAATAGAGACGA- TCCGAACT	GCGCAGAAGGCA- GTGTTT

Primer name	Forward primer (5'-3')	Reverse primer (5'-3')
CACS	TGGGAAGATTCTTA- TGAAGTGC	CTCGTCAAATTT- ACACATTGGT
PP2A	CAACAGGTGATATT- GGATTATGAT	GCCAGCTCATCC- TCATATAAG
AAT4	TTCGTACACGCAAC- GAGA	TGAAGAGGGTAA- GGCTTG
NAAT1	GAGGAGCTGTGAAA- GGAGCA	CCCTCCACGACA- GTTCCAAT
AAT9	CAGAAACAGCGAAC- CAGAGC	AACTTCATCCCCA- CCGAGTT
AAT14	GGGAATAGAGACGA- TCCGAACT	GCGCAGAAGGCA- GTGTTT

Gene	Start position	End position	Orientation	Gene	Start position	End position	Orientation
CucsaAAT1	5496646	5495195	Reverse	CucsaAAT9	5654159	5656355	Forward
CucsaAAT2	5583941	5582553	Reverse	CucsaAAT10	5681664	5683484	Forward
CucsaAAT3	5590531	5589140	Reverse	CucsaAAT11	5691672	5690257	Reverse
CucsaAAT4	5596447	5595167	Reverse	CucsaAAT12	7077986	7075008	Reverse
CucsaAAT5	5605793	5607184	Forward	CucsaAAT13	7083801	7085207	Forward
CucsaAAT6	5619299	5621364	Forward	CucsaAAT14	7090825	7094963	Forward
CucsaAAT7	5630016	5631894	Forward	CucsaNAAT1	5648115	5646235	Reverse
CucsaAAT8	5636899	5638943	Forward	CucsaNAAT2	5670107	5668263	Reverse

Gene	Start position	End position	Orientation	Gene	Start position	End position	Orientation
CucsaAAT1	5496646	5495195	Reverse	CucsaAAT9	5654159	5656355	Forward
CucsaAAT2	5583941	5582553	Reverse	CucsaAAT10	5681664	5683484	Forward
CucsaAAT3	5590531	5589140	Reverse	CucsaAAT11	5691672	5690257	Reverse
CucsaAAT4	5596447	5595167	Reverse	CucsaAAT12	7077986	7075008	Reverse
CucsaAAT5	5605793	5607184	Forward	CucsaAAT13	7083801	7085207	Forward
CucsaAAT6	5619299	5621364	Forward	CucsaAAT14	7090825	7094963	Forward
CucsaAAT7	5630016	5631894	Forward	CucsaNAAT1	5648115	5646235	Reverse
CucsaAAT8	5636899	5638943	Forward	CucsaNAAT2	5670107	5668263	Reverse

Protein	Domain architecture	No. of amino acids	Protein size (kDa)	Theoretical pI
CucsaAAT1	AAT	484	54.9	8.3
CucsaAAT2	AAT	463	53.6	7.9
CucsaAAT3	AAT	463	53.5	8.0
CucsaAAT4	AAT	466	53.7	8.2
CucsaAAT5	AAT	463	53.3	7.5
CucsaAAT6	AAT	464	53.4	7.5
CucsaAAT7	AAT	471	53.5	5.1
CucsaAAT8	AAT	475	54.7	8.3
CucsaAAT9	AAT	474	55.0	6.7
CucsaAAT10	AAT	476	54.9	5.7
CucsaAAT11	AAT	482	55.4	7.0
CucsaAAT12	AAT	502	57.6	9.0
CucsaAAT13	AAT	468	53.9	7.7
CucsaAAT14	AAT	505	58.2	6.8
CucsaNAAT1	NAAT	626	70.4	5.1
CucsaNAAT2	NAAT	614	69.4	4.9