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.

References

AbubakarAS, FengX, GaoG, ChunmingY, ChenJ, ChenK, WangX, MouP, ShaoD, ChenP, ZhuA. Genome wide characterization of R2R3 MYB transcription factor from Apocynum venetum revealed potential stress tolerance and flavonoid biosynthesis genes. Genomics, 2022, 114(2): 110275

AugustyniakA, PerlikowskiD, RapaczM, KoscielniakJ, KosmalaA. Insight into cellular proteome of Lolium multiflorum/Festuca arundinacea introgression forms to decipher crucial mechanisms of cold acclimation in forage grasses. Plant Sci, 2018, 272: 22-31

BanoH, AtharHUR, ZafarZU, OgbagaCC, AshrafM. 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, 2021, 172(2): 603-614

BowmanJL, EshedY, BaumSF. Establishment of polarity in angiosperm lateral organs. Trends Genet TIG, 2002, 18(3): 134-141

CaiG, XuY, ZhangS, ChenT, LiuG, LiZ, ZhuY, WangG. A tomato chloroplast-targeted DnaJ protein, SlDnaJ20 maintains the stability of photosystem I/II under chilling stress. Plant Signal Behav, 2022

ChenK, AroraR. 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, 2014, 150(3): 374-387

ChenK, RenautJ, SergeantK, WeiH, AroraR. Proteomic changes associated with freeze-thaw injury and post-thaw recovery in onion (Allium cepa L.) scales. Plant Cell Environ, 2013, 36(4): 892-905

ChenY, ChenZ, KangJ, KangD, GuH, QinG. AtMYB14 Regulates Cold Tolerance in Arabidopsis. Plant Mol Biol Report, 2013, 31(1): 87-97

ChengL, LiX, HuangX, MaT, LiangY, MaX, PengX, JiaJ, ChenS, ChenY, DengB, LiuG. Overexpression of Leymus chinensis R1-MYB transcription factor LcMYB1 confers salt tolerance in transgenic Arabidopsis. Plant Physiol Biochem, 2013, 70: 252-260

CuiX, ZhangY, QiF, GaoJ, ChenY, ZhangC. Overexpression of a moso bamboo (Phyllostachys edulis) transcription factor gene PheWRKY1 enhances disease resistance in transgenic Arabidopsis thaliana. Botany, 2013, 91(7): 486-494

DaiL, ZhengT, LiuC, LiK, QuG. Construction of plant Expression vector and genetic transformation analysis of Arabidopsis thaliana CYCD2;1 gene in Nicotiana tabacum. Bullet Botanical Res, 2016, 36(2): 283-290

DaiL, FengZ, PanX, XuY, LiP, LefohnAS, HarmensH, KobayashiK. Increase of apoplastic ascorbate induced by ozone is insufficient to remove the negative effects in tobacco, soybean and poplar. Environ Pollut, 2019, 245: 380-388

DengY, KongF, ZhouB, ZhangS, YueM, MengQ. Heterology expression of the tomato LeLhcb2 gene confers elevated tolerance to chilling stress in transgenic tobacco. Plant Physiol Biochem, 2014, 80: 318-327

DingF, WangM, LiuB, ZhangS. Exogenous melatonin mitigates photoinhibition by accelerating non-photochemical quenching in tomato seedlings exposed to moderate light during chilling. Front Plant Sci, 2017

FoyerCH, RubanAV, NoctorG. Viewing oxidative stress through the lens of oxidative signalling rather than damage. Biochem J, 2017, 474: 877-883

GerardoLC, BasilioHJ, OsunaET, AdrianaSBJ, LopezMM, AlbertoLRL, LeonFJ. Alterations in photosynthetic processes due to the effect of oxidative stress induced by cold and UV-B radiation. Int J Agric Biol, 2017, 19(6): 1363-1371

GuptaAS, HeinenJL, HoladayAS, BurkeJJ, AllenRD. Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase. Proc Natl Acad Sci USA, 1993, 90(4): 1629-1633

HanX, LiP, ZouL, TanW, ZhengT, ZhangD, LinH. GOLDEN2-LIKE transcription factors coordinate the tolerance to Cucumber mosaic virus in Arabidopsis. Biochem Biophys Res Commun, 2016, 477(4): 626-632

HuenerNPA, BodeR, DahalK, HollisL, RossoD, KrolM, IvanovAG. Chloroplast redox imbalance governs phenotypic plasticity: the “grand design of photosynthesis” revisited. Front Plant Sci, 2012

ItoY, KatsuraK, MaruyamaK, TajiT, KobayashiM, SekiM, ShinozakiK, YamaguchiSK. Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol, 2006, 47(1): 141-153

JiangC, ZhengQ, LiuZ, XuW, LiuL, ZhaoG, LongX. Overexpression of Arabidopsis thaliana Na+/H+ antiporter gene enhanced salt resistance in transgenic poplar (Populus x euramericana ‘Neva’). Trees-Struct Function, 2012, 26(3): 685-694

KongF, DengY, ZhouB, WangG, WangY, MengQ. A chloroplast-targeted DnaJ protein contributes to maintenance of photosystem II under chilling stress. J Exp Bot, 2014, 65(1): 143-158

KromdijkJ, GlowackaK, LeonelliL, GabillyST, IwaiM, NiyogiKK, LongSP. Improving photosynthesis and crop productivity by accelerating recovery from photoprotection. Science, 2016, 354(6314): 857-861

LiX, YangW, LiuS, LiX, JiaJ, ZhaoP, ChengL, QiD, ChenS, LiuG. LcFIN2, a novel chloroplast protein gene from Leymus chinensis, enhances tolerance to low temperature in Arabidopsis and rice. Physiol Plant, 2019, 166(2): 628-645

LiB, WangX, WangX, XiZ. An AP2/ERF transcription factor VvERF63 positively regulates cold tolerance in Arabidopsis and grape leaves. Environ Exp Botany, 2023

LinK, HwangW, LoH. Chilling stress and chilling tolerance of sweet potato as sensed by chlorophyll fluorescence. Photosynthetica, 2007, 45(4): 628-632

LiuQ, KasugaM, SakumaY, AbeH, MiuraS, YamaguchiSK, ShinozakiK. 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, 1998, 10(8): 1391-1406

LiuY, HanX, ZhanX, YangJ, WangY, SongQ, ChenX. Regulation of calcium on peanut photosynthesis under low night temperature stress. J Integr Agric, 2013, 12(12): 2172-2178

LiuG, LiF, ShiG, WangL, WangL, FanL. Identification of MADS-Box transcription factors in iris laevigata and functional assessment of IlSEP3 and IlSVP during flowering. Int J Mol Sci, 2022

LiuX, XiaW, ZhangX, LiA, QinJ, SunH, LiJ, ZhuJ. Overexpression of the SiLEA5 Gene in Saussurea involucrata increases the low-temperature tolerance of transgenic tomatoes. Horticulturae, 2022

LouY, SunH, ZhuC, YangK, LiX, GaoZ. PeVDE, a violaxanthin de-epoxidase gene from moso bamboo, confers photoprotection ability in transgenic Arabidopsis under high light. Fron Plant Sci, 2022

MalnoeA, SchultinkA, ShahrasbiS, RumeauD, HavauxM, NiyogiKK. The plastid lipocalin LCNP Is required for sustained photoprotective energy dissipation in Arabidopsis. Plant Cell, 2018, 30(1): 196-208

MattilaH, TyystjarviE. Light-induced damage to photosystem II at a very low temperature (195 K) depends on singlet oxygen. Physiol Plant, 2022

MukherjeeS, MukherjeeA, DasP, BandyopadhyayS, ChattopadhyayD, ChatterjeeJ, MajumderAL. 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, 2021, 145(3): 561-578

MurchieEH, RubanAV. Dynamic non-photochemical quenching in plants: from molecular mechanism to productivity. Plant J, 2020, 101(4): 885-896

SantariusKA. Freezing of isolated thylakoid membranes in complex media. V. Inactivation and protection of electron transport reactions. Photosynthesis Res, 1990, 23(1): 49-58

SongX, DiaoJ, JiJ, WangG, LiZ, WuJ, JosineTL, WangY. Overexpression of lycopene epsilon-cyclase gene from Lycium chinense confers tolerance to chilling stress in Arabidopsis thaliana. Gene, 2016, 576(1): 395-403

SongQ, LiuY, PangJ, YongJWH, ChenY, BaiC, GilleC, ShiQ, WuD, HanX, LiT, SiddiqueKHM, LambersH. Supplementary calcium restores peanut (Arachis hypogaea) growth and photosynthetic capacity under low nocturnal temperature. Front Plant Sci, 2020

SongQ, WangX, LiuY, BresticM, YangX. StLTO1, a lumen thiol oxidoreductase in Solanum tuberosum L., enhances the cold resistance of potato plants. Plant Sci, 2022

SunW, WeiJ, WuG, XuH, ChenY, YaoM, ZhanJ, YanJ, WuN, ChenH, BuT, TangZ, LiQ. CqZF-HD14 enhances drought tolerance in quinoa seedlings through interaction with CqHIPP34 and CqNAC79. Plant Sci, 2022

TajZ, ChallabathulaD. 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, 2021

WangY, ShiM, ZhangX, LiuY, XueF, SunJ, LiY. Cloning and expression analysis of GhMYB113 Gene in Gossypium hirsutum. Acta Botan Boreali Occiden Sin, 2013, 33(5): 878-884

WangF, NanW, ZhangL, AhammedGJ, ChenX, XiangX, ZhouJ, XiaX, ShiK, JingquanY, FoyerCH, ZhouY. Light signaling-dependent regulation of photoinhibition and photoprotection in tomato. Plant Physiol, 2018, 176(2): 1311-1326

WangY, ChenQ, ZhengJ, ZhangZ, GaoT, LiC, MaF. Overexpression of the tyrosine decarboxylase gene MdTyDC in apple enhances long-term moderate drought tolerance and WUE. Plant Sci, 2021

WeiY, ChenH, WangL, ZhaoQ, WangD. Zhang T (2022) Cold acclimation alleviates cold stress-induced PSII inhibition and oxidative damage in tobacco leaves. Plant Signal Behav, 2021, 10(1080/15592324): 2013638

XieY, ChenP, YanY, BaoC, LiX, WangL, ShenX, LiH, LiuX, NiuC, ZhuC, FangN, ShaoY, ZhaoT, YuJ, ZhuJ, XuL, NockerS, MaF, GuanQ. An atypical R2R3 MYB transcription factor increases cold hardiness by CBF-dependent and CBF-independent pathways in apple. New Phytol, 2018, 218(1): 201-218

XuZ. Rothstein SJ (2018) ROS-Induced anthocyanin production provides feedback protection by scavenging ROS and maintaining photosynthetic capacity in Arabidopsis. Plant Signal Behav, 2018, 10(1080/15592324): 1451708

YamaguchiSK, ShinozakiK. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol, 2006, 57(1): 781-803

YangJ, YuS, ShiG, YanL, LvR, MaZ, WangL. Comparative analysis of R2R3-MYB transcription factors in the flower of Iris laevigata identifies a novel gene regulating tobacco cold tolerance. Plant Biol J, 2022, 24: 1066-1075

ZengJ, HuW, HuX, TaoH, ZhongL, LiuL. Upregulation of the mitochondrial alternative oxidase pathway improves PSII function and photosynthetic electron transport in tomato seedlings under chilling stress. Photosynthetica, 2022, 60(2): 271-279

ZhanX, QiJ, ShenQ, HeB, MaoB. Regulation of phenylpropanoid metabolism during moderate freezing and post-freezing recovery in Dendrobium offcinale. J Plant Interact, 2022, 17(1): 290-300

ZhangY, YangJ, GuoS, MengJ, ZhangY, WanS, HeQ, LiX. Over-expression of the Arabidopsis CBF1 gene improves resistance of tomato leaves to low temperature under low irradiance. Plant Biol, 2011, 13(2): 362-367

ZhangZ, JiaY, GaoH, ZhangL, LiH, MengQ. Characterization of PSI recovery after chilling-induced photoinhibition in cucumber (Cucumis sativus L.) leaves. Planta, 2011, 234(5): 883-889

ZhangH, YafanH, BaoG, CuiX, ZhangJ. VaMYB44 transcription factor from Chinese wild Vitis amurensis negatively regulates cold tolerance in transgenic Arabidopsis thaliana and V. vinifera. Plant Cell Rep, 2022, 41(8): 1673-1691

ZhangL, SongJ, LinR, TangM, ShaoS, YuJ, ZhouY. Tomato SlMYB15 transcription factor targeted by sly-miR156e-3p positively regulates ABA-mediated cold tolerance. J Exp Bot, 2022, 73(22): 7538-7551

ZhaoH, AbulaiziA, WangC, LanH. Overexpression of CgbHLH001, a positive regulator to adversity, enhances the photosynthetic capacity of maize seedlings under drought stress. Agronomy, 2022, 12(5): 1149

ZhuangK, KongF, ZhangS, MengC, YangM, LiuZ, WangY, MaN, MengQ. Whirly1 enhances tolerance to chilling stress in tomato via protection of photosystem II and regulation of starch degradation. New Phytol, 2019, 221(4): 1998-2012