Over-expression of the Iris laevigata cold-resistance gene MYB97 improves photosynthetic capacity and photoprotection in tobacco (Nicotiana tabacum)

Abstract

In northern China, light and temperature are major limiting factors for plant growth, particularly during seed production and seedling establishment. While previous studies suggested a possible role for the MYB97 gene in cold-stress, confirmation through documented evidence was lacking. In this study, we transformed the MYB97 gene from Iris laevigata into tobacco, and discovered that the gene boosted photosynthesis, photoprotection and resilience to cold. The transgenic tobacco seeds exhibited enhanced germination and accelerated seedling growth. Moreover, these plants had decreased levels of MDA (Malondialdehyde) and relative conductance, coupled with elevated concentrations of proline and soluble sugars. This response was accompanied by heightened activity of antioxidant enzymes during periods of cold stress (4 and − 2 °C). Exposure to low temperatures (0–15 °C) also reduced heights but accentuated primary root growth in transgenic tobacco plants. Additionally, tobacco leaves showed an increased growth along with higher chlorophyll levels, net photosynthetic rates, stomatal conductance, transpiration rates and non-photochemical quenching coefficient. This study shows that IlMYB97 (The MYB97 genes in I. laevigata) improves cold-resistance, and enhances photosynthesis and photoprotective ability, and thus overall growth and development. These findings would offer the genetic resources to further study cold resistance and photosynthesis.

Keywords

Iris laevigata / MYB97 / Photosynthesis / Photoprotection / Cold stress

References

[1]	Abubakar AS, Feng X, Gao G, Chunming Y, Chen J, Chen K, Wang X, Mou P, Shao D, Chen P, Zhu A (2022) Genome wide characterization of R2R3 MYB transcription factor from Apocynum venetum revealed potential stress tolerance and flavonoid biosynthesis genes. Genomics 114(2):110275. https://doi.org/10.1016/j.ygeno.2022.110275
[2]	Augustyniak A, Perlikowski D, Rapacz M, Koscielniak J, Kosmala A (2018) Insight into cellular proteome of Lolium multiflorum/Festuca arundinacea introgression forms to decipher crucial mechanisms of cold acclimation in forage grasses. Plant Sci 272:22–31. https://doi.org/10.1016/j.plantsci.2018.04.002
[3]	Bano H, Athar HUR, Zafar ZU, Ogbaga CC, Ashraf M (2021) Peroxidase activity and operation of photo-protective component of NPQ play key roles in drought tolerance of mung bean Vigna radiata (L.) Wilcziek. Physiol Plant 172(2):603–614. https://doi.org/10.1111/ppl.13337
[4]	Bowman JL, Eshed Y, Baum SF (2002) Establishment of polarity in angiosperm lateral organs. Trends Genet TIG 18(3):134–141. https://doi.org/10.1016/S0168-9525(01)02601-4
[5]	Cai G, Xu Y, Zhang S, Chen T, Liu G, Li Z, Zhu Y, Wang G (2022) A tomato chloroplast-targeted DnaJ protein, SlDnaJ20 maintains the stability of photosystem I/II under chilling stress. Plant Signal Behav. https://doi.org/10.1080/155923242139116
[6]	Chen K, Arora R (2014) Understanding the cellular mechanism of recovery from freeze-thaw injury in spinach: possible role of aquaporins, heat shock proteins, dehydrin and antioxidant system. Physiol Plant 150(3):374–387. https://doi.org/10.1111/ppl.12090
[7]	Chen K, Renaut J, Sergeant K, Wei H, Arora R (2013a) Proteomic changes associated with freeze-thaw injury and post-thaw recovery in onion (Allium cepa L.) scales. Plant Cell Environ 36(4):892–905. https://doi.org/10.1111/pce.12027
[8]	Chen Y, Chen Z, Kang J, Kang D, Gu H, Qin G (2013b) AtMYB14 Regulates Cold Tolerance in Arabidopsis. Plant Mol Biol Report 31(1):87–97. https://doi.org/10.1007/s11105-012-0481-z
[9]	Cheng L, Li X, Huang X, Ma T, Liang Y, Ma X, Peng X, Jia J, Chen S, Chen Y, Deng B, Liu G (2013) Overexpression of Leymus chinensis R1-MYB transcription factor LcMYB1 confers salt tolerance in transgenic Arabidopsis. Plant Physiol Biochem 70:252–260. https://doi.org/10.1016/j.plaphy.2013.05.025
[10]	Cui X, Zhang Y, Qi F, Gao J, Chen Y, Zhang C (2013) Overexpression of a moso bamboo (Phyllostachys edulis) transcription factor gene PheWRKY1 enhances disease resistance in transgenic Arabidopsis thaliana. Botany 91(7):486–494. https://doi.org/10.1139/cjb-2012-0219
[11]	Dai L, Zheng T, Liu C, Li K, Qu G (2016) Construction of plant Expression vector and genetic transformation analysis of Arabidopsis thaliana CYCD2;1 gene in Nicotiana tabacum. Bullet Botanical Res 36(2):283–290
[12]	Dai L, Feng Z, Pan X, Xu Y, Li P, Lefohn AS, Harmens H, Kobayashi K (2019) Increase of apoplastic ascorbate induced by ozone is insufficient to remove the negative effects in tobacco, soybean and poplar. Environ Pollut 245:380–388. https://doi.org/10.1016/j.envpol.2018.11.030
[13]	Deng Y, Kong F, Zhou B, Zhang S, Yue M, Meng Q (2014) Heterology expression of the tomato LeLhcb2 gene confers elevated tolerance to chilling stress in transgenic tobacco. Plant Physiol Biochem 80:318–327. https://doi.org/10.1016/j.plaphy.2014.04.017
[14]	Ding F, Wang M, Liu B, Zhang S (2017) Exogenous melatonin mitigates photoinhibition by accelerating non-photochemical quenching in tomato seedlings exposed to moderate light during chilling. Front Plant Sci. https://doi.org/10.3389/fpls.2017.00244
[15]	Foyer CH, Ruban AV, Noctor G (2017) Viewing oxidative stress through the lens of oxidative signalling rather than damage. Biochem J 474:877–883. https://doi.org/10.1042/BCJ20160814
[16]	Gerardo LC, Basilio HJ, Osuna ET, Adriana SBJ, Lopez MM, Alberto LRL, Leon FJ (2017) Alterations in photosynthetic processes due to the effect of oxidative stress induced by cold and UV-B radiation. Int J Agric Biol 19(6):1363–1371. https://doi.org/10.17957/IJAB/15.0418
[17]	Gupta AS, Heinen JL, Holaday AS, Burke JJ, Allen RD (1993) Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase. Proc Natl Acad Sci USA 90(4):1629–1633. https://doi.org/10.1073/as.90.4.1629
[18]	Han X, Li P, Zou L, Tan W, Zheng T, Zhang D, Lin H (2016) GOLDEN2-LIKE transcription factors coordinate the tolerance to Cucumber mosaic virus in Arabidopsis. Biochem Biophys Res Commun 477(4):626–632. https://doi.org/10.1016/j.bbrc.2016.06.110
[19]	Huener NPA, Bode R, Dahal K, Hollis L, Rosso D, Krol M, Ivanov AG (2012) Chloroplast redox imbalance governs phenotypic plasticity: the “grand design of photosynthesis” revisited. Front Plant Sci. https://doi.org/10.3389/fpls.2012.00255
[20]	Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi SK (2006) Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol 47(1):141–153. https://doi.org/10.1093/pcp/pci230
[21]	Jiang C, Zheng Q, Liu Z, Xu W, Liu L, Zhao G, Long X (2012) Overexpression of Arabidopsis thaliana Na+/H+ antiporter gene enhanced salt resistance in transgenic poplar (Populus x euramericana ‘Neva’). Trees-Struct Function 26(3):685–694. https://doi.org/10.1007/s00468-011-0635-x
[22]	Kong F, Deng Y, Zhou B, Wang G, Wang Y, Meng Q (2014) A chloroplast-targeted DnaJ protein contributes to maintenance of photosystem II under chilling stress. J Exp Bot 65(1):143–158. https://doi.org/10.1093/jxb/ert357
[23]	Kromdijk J, Glowacka K, Leonelli L, Gabilly ST, Iwai M, Niyogi KK, Long SP (2016) Improving photosynthesis and crop productivity by accelerating recovery from photoprotection. Science 354(6314):857–861. https://doi.org/10.1126/science.aai8878
[24]	Li X, Yang W, Liu S, Li X, Jia J, Zhao P, Cheng L, Qi D, Chen S, Liu G (2019) LcFIN2, a novel chloroplast protein gene from Leymus chinensis, enhances tolerance to low temperature in Arabidopsis and rice. Physiol Plant 166(2):628–645. https://doi.org/10.1111/ppl.12811
[25]	Li B, Wang X, Wang X, Xi Z (2023) An AP2/ERF transcription factor VvERF63 positively regulates cold tolerance in Arabidopsis and grape leaves. Environ Exp Botany. https://doi.org/10.1016/j.envexpbot.2022.105124
[26]	Lin K, Hwang W, Lo H (2007) Chilling stress and chilling tolerance of sweet potato as sensed by chlorophyll fluorescence. Photosynthetica 45(4):628–632. https://doi.org/10.1007/s11099-007-0108-z
[27]	Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi SK, Shinozaki K (1998) Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively. Arabidopsis Plant Cell 10(8):1391–1406. https://doi.org/10.2307/3870648
[28]	Liu Y, Han X, Zhan X, Yang J, Wang Y, Song Q, Chen X (2013) Regulation of calcium on peanut photosynthesis under low night temperature stress. J Integr Agric 12(12):2172–2178. https://doi.org/10.1016/S2095-3119(13)60411-6
[29]	Liu G, Li F, Shi G, Wang L, Wang L, Fan L (2022a) Identification of MADS-Box transcription factors in iris laevigata and functional assessment of IlSEP3 and IlSVP during flowering. Int J Mol Sci. https://doi.org/10.3390/ijms23179950
[30]	Liu X, Xia W, Zhang X, Li A, Qin J, Sun H, Li J, Zhu J (2022b) Overexpression of the SiLEA5 Gene in Saussurea involucrata increases the low-temperature tolerance of transgenic tomatoes. Horticulturae. https://doi.org/10.3390/horticulturae8111023
[31]	Lou Y, Sun H, Zhu C, Yang K, Li X, Gao Z (2022) PeVDE, a violaxanthin de-epoxidase gene from moso bamboo, confers photoprotection ability in transgenic Arabidopsis under high light. Fron Plant Sci. https://doi.org/10.3389/fpls.2022.927949
[32]	Malnoe A, Schultink A, Shahrasbi S, Rumeau D, Havaux M, Niyogi KK (2018) The plastid lipocalin LCNP Is required for sustained photoprotective energy dissipation in Arabidopsis. Plant Cell 30(1):196–208. https://doi.org/10.1105/tpc.17.00536
[33]	Mattila H, Tyystjarvi E (2022) Light-induced damage to photosystem II at a very low temperature (195 K) depends on singlet oxygen. Physiol Plant. https://doi.org/10.1111/ppl.13824
[34]	Mukherjee S, Mukherjee A, Das P, Bandyopadhyay S, Chattopadhyay D, Chatterjee J, Majumder AL (2021) A salt-tolerant chloroplastic FBPase from Oryza coarctata confers improved photosynthesis with higher yield and multi-stress tolerance to indica rice. Plant Cell Tissue Organ Cult 145(3):561–578. https://doi.org/10.1007/s11240-021-02026-1
[35]	Murchie EH, Ruban AV (2020) Dynamic non-photochemical quenching in plants: from molecular mechanism to productivity. Plant J 101(4):885–896. https://doi.org/10.1111/tpj.14601
[36]	Santarius KA (1990) Freezing of isolated thylakoid membranes in complex media. V. Inactivation and protection of electron transport reactions. Photosynthesis Res 23(1):49–58. https://doi.org/10.1007/BF00030062
[37]	Song X, Diao J, Ji J, Wang G, Li Z, Wu J, Josine TL, Wang Y (2016) Overexpression of lycopene epsilon-cyclase gene from Lycium chinense confers tolerance to chilling stress in Arabidopsis thaliana. Gene 576(1):395–403. https://doi.org/10.1016/j.gene.2015.10.051
[38]	Song Q, Liu Y, Pang J, Yong JWH, Chen Y, Bai C, Gille C, Shi Q, Wu D, Han X, Li T, Siddique KHM, Lambers H (2020) Supplementary calcium restores peanut (Arachis hypogaea) growth and photosynthetic capacity under low nocturnal temperature. Front Plant Sci. https://doi.org/10.3389/fpls.2019.01637
[39]	Song Q, Wang X, Liu Y, Brestic M, Yang X (2022) StLTO1, a lumen thiol oxidoreductase in Solanum tuberosum L., enhances the cold resistance of potato plants. Plant Sci. https://doi.org/10.1016/j.plantsci.2022.111481
[40]	Sun W, Wei J, Wu G, Xu H, Chen Y, Yao M, Zhan J, Yan J, Wu N, Chen H, Bu T, Tang Z, Li Q (2022) CqZF-HD14 enhances drought tolerance in quinoa seedlings through interaction with CqHIPP34 and CqNAC79. Plant Sci. https://doi.org/10.1016/j.plantsci.2022.111406
[41]	Taj Z, Challabathula D (2021) Protection of photosynthesis by halotolerant staphylococcus sciuri ET101 in tomato (Lycoperiscon esculentum) and rice (Oryza sativa) plants during salinity stress: possible interplay between carboxylation and oxygenation in stress mitigation. Front Microbiol. https://doi.org/10.3389/fmicb.2020.547750
[42]	Wang Y, Shi M, Zhang X, Liu Y, Xue F, Sun J, Li Y (2013) Cloning and expression analysis of GhMYB113 Gene in Gossypium hirsutum. Acta Botan Boreali Occiden Sin 33(5):878–884
[43]	Wang F, Nan W, Zhang L, Ahammed GJ, Chen X, Xiang X, Zhou J, Xia X, Shi K, Jingquan Y, Foyer CH, Zhou Y (2018) Light signaling-dependent regulation of photoinhibition and photoprotection in tomato. Plant Physiol 176(2):1311–1326. https://doi.org/10.1104/pp.17.01143
[44]	Wang Y, Chen Q, Zheng J, Zhang Z, Gao T, Li C, Ma F (2021) Overexpression of the tyrosine decarboxylase gene MdTyDC in apple enhances long-term moderate drought tolerance and WUE. Plant Sci. https://doi.org/10.1016/j.plantsci.2021.111064
[45]	Wei Y, Chen H, Wang L, Zhao Q, Wang D (2021) Zhang T (2022) Cold acclimation alleviates cold stress-induced PSII inhibition and oxidative damage in tobacco leaves. Plant Signal Behav 10(1080/15592324):2013638
[46]	Xie Y, Chen P, Yan Y, Bao C, Li X, Wang L, Shen X, Li H, Liu X, Niu C, Zhu C, Fang N, Shao Y, Zhao T, Yu J, Zhu J, Xu L, Nocker S, Ma F, Guan Q (2018) An atypical R2R3 MYB transcription factor increases cold hardiness by CBF-dependent and CBF-independent pathways in apple. New Phytol 218(1):201–218. https://doi.org/10.1111/nph.14952
[47]	Xu Z (2018) Rothstein SJ (2018) ROS-Induced anthocyanin production provides feedback protection by scavenging ROS and maintaining photosynthetic capacity in Arabidopsis. Plant Signal Behav 10(1080/15592324):1451708
[48]	Yamaguchi SK, Shinozaki K (2006) Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol 57(1):781–803. https://doi.org/10.1146/annurev.arplant.57.032905.105444
[49]	Yang J, Yu S, Shi G, Yan L, Lv R, Ma Z, Wang L (2022) Comparative analysis of R2R3-MYB transcription factors in the flower of Iris laevigata identifies a novel gene regulating tobacco cold tolerance. Plant Biol J 24:1066–1075. https://doi.org/10.1111/plb.13452
[50]	Zeng J, Hu W, Hu X, Tao H, Zhong L, Liu L (2022) Upregulation of the mitochondrial alternative oxidase pathway improves PSII function and photosynthetic electron transport in tomato seedlings under chilling stress. Photosynthetica 60(2):271–279. https://doi.org/10.32615/ps.2022.019
[51]	Zhan X, Qi J, Shen Q, He B, Mao B (2022) Regulation of phenylpropanoid metabolism during moderate freezing and post-freezing recovery in Dendrobium offcinale. J Plant Interact 17(1):290–300. https://doi.org/10.1080/17429145.2022.2036377
[52]	Zhang Y, Yang J, Guo S, Meng J, Zhang Y, Wan S, He Q, Li X (2011a) Over-expression of the Arabidopsis CBF1 gene improves resistance of tomato leaves to low temperature under low irradiance. Plant Biol 13(2):362–367. https://doi.org/10.1111/j.1438-8677.2010.00365.x
[53]	Zhang Z, Jia Y, Gao H, Zhang L, Li H, Meng Q (2011b) Characterization of PSI recovery after chilling-induced photoinhibition in cucumber (Cucumis sativus L.) leaves. Planta 234(5):883–889. https://doi.org/10.1007/s00425-011-1447-3
[54]	Zhang H, Yafan H, Bao G, Cui X, Zhang J (2022a) VaMYB44 transcription factor from Chinese wild Vitis amurensis negatively regulates cold tolerance in transgenic Arabidopsis thaliana and V. vinifera. Plant Cell Rep 41(8):1673–1691. https://doi.org/10.1007/s00299-022-02883-w
[55]	Zhang L, Song J, Lin R, Tang M, Shao S, Yu J, Zhou Y (2022b) Tomato SlMYB15 transcription factor targeted by sly-miR156e-3p positively regulates ABA-mediated cold tolerance. J Exp Bot 73(22):7538–7551. https://doi.org/10.1093/jxb/erac370
[56]	Zhao H, Abulaizi A, Wang C, Lan H (2022) Overexpression of CgbHLH001, a positive regulator to adversity, enhances the photosynthetic capacity of maize seedlings under drought stress. Agronomy 12(5):1149. https://doi.org/10.3390/agronomy12051149
[57]	Zhuang K, Kong F, Zhang S, Meng C, Yang M, Liu Z, Wang Y, Ma N, Meng Q (2019) Whirly1 enhances tolerance to chilling stress in tomato via protection of photosystem II and regulation of starch degradation. New Phytol 221(4):1998–2012. https://doi.org/10.1111/nph.15532