Identification and functional characterization of safflower cysteine protease 1 as negative regulator in response to low-temperature stress in transgenic Arabidopsis

Hong, Yingqi; Lv, Yuanda; Zhang, Jianyi; Ahmad, Naveed; Li, Youbao; Wang, Nan; Liu, Xiuming; Yao, Na; Li, Xiaokun

doi:10.1007/s00425-022-03875-6

Cited by 9 publications

(4 citation statements)

References 58 publications

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“…Loss of function of CaCP using the virus-induced gene-silencing technique led to enhanced tolerance to salt-induced and osmotic-induced stress [32]. Transgenic plants inhibited by CtCP1-anti showed higher tolerance to low temperature than WT and CtCP1-OE plants [35]. The overexpression of sweet potato SPCP3 in Arabidopsis conferred sensitivity to drought stress [36].…”

Section: Discussionmentioning

confidence: 99%

“…The mixture contained 50 mM potassium phosphate buffer (pH 7.0), 0.1 mM EDTA, 50 mM guaiacol, 2% (v/v) H 2 O 2 , and 0.1 mL of the enzyme extract. Changes in the absorbance were measured at 470 nm [35]. The catalase (CAT) activity was analyzed in 3 mL reaction mixture (50 mM potassium phosphate buffer (pH 7.0), 2 mM Na 2 -EDTA, 0.1 mol/L H 2 O 2 and 0.2 mL of enzyme extract), and the changes in the absorbance at 240 nm were measured over a period of 1 min.…”

Section: Determination Of Physiological and Biochemical Activitiesmentioning

confidence: 99%

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A Novel Senescence-Specific Gene (ZmSAG39) Negatively Regulates Darkness and Drought Responses in Maize

Wang

Gao

Chen

et al. 2022

IJMS

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The papain-like cysteine proteases (PLCPs) is a subfamily of cysteine proteases that plays an important role in leaf senescence, and some of its members are involved in the regulation of plant growth and development under stress. In this study, we cloned a new gene, ZmSAG39, from maize. Expression profile analysis showed that ZmSAG39 was induced by darkness and drought treatments. In addition, the ZmSAG39 overexpression in maize accelerated the senescence of maize leaves under darkness and drought treatments. However, the knockout of ZmSAG39 in maize enhanced the resistance of maize to darkness and drought stresses and reduced the degree of senescence of maize leaves. Under drought stress, compared with WT plants, the knockout lines had a higher seed germination rate, seedling survival rate and chlorophyll content, and lower reactive oxygen species (ROS) level and malondialdehyde (MDA) content. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that ZmSAG39 negatively regulated some stress-related genes but positively regulated senescence-related genes under darkness and drought stress conditions. To summarize, these results indicate that ZmSAG39 is a senescence-related gene and plays a negative role in response to darkness and drought stresses. This study laid a theoretical foundation for the innovation of maize germplasm resources with high quality, high yield and strong stress resistance.

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Section: Discussionmentioning

confidence: 99%

Section: Determination Of Physiological and Biochemical Activitiesmentioning

confidence: 99%

A Novel Senescence-Specific Gene (ZmSAG39) Negatively Regulates Darkness and Drought Responses in Maize

Wang

Gao

Chen

et al. 2022

IJMS

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“…Meanwhile, functions of some genes have been identified through heterologous expression in Arabidopsis . For instance, safflower chalcone isomerase (CHI) [ 3 ], cytochrome P450 (CYP) [ 4 ], and cysteine protease 1 (CP1) [ 5 ] have been functionally characterized using Arabidopsis models . However, many medicinal plants, such as safflower, contain unique secondary metabolites.…”

Section: Introductionmentioning

confidence: 99%

The establishment of transient expression systems and their application for gene function analysis of flavonoid biosynthesis in Carthamus tinctorius L

Xian

Ren

et al. 2023

BMC Plant Biol

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Background Safflower (Carthamus tinctorius L.) is an important economic crop and a traditional medicinal material rich in flavonoids, which can alleviate cardiovascular and cerebrovascular pathologies. Thus, many candidate genes involved in safflower flavonoid biosynthesis have been cloned. However, owing to the lack of a homologous gene expression system, research on gene function is limited to model plants. Therefore, a gene function identification protocol for safflower must be established. Results In the present study, using safflower callus as the experimental material, Agrobacterium and biolistic transient expression systems were established. In the Agrobacterium transient expression system, the highest transformation rate was obtained at the original Agrobacterium concentration of OD600 0.4, infiltration concentration of OD600 0.6, infection for 20 min, co-culture for 3 days, and acetosyringone concentration of 100 μmol·L−1. In the biolistic transient expression system, the highest transformation efficiency was observed at helium pressure of 1,350 psi, vacuum degree of -0.8 bar, flight distance of 6.5 cm, one round of bombardment, plasmid concentration of 3 μg·shot−1, and gold particle concentration of 100 μg·shot−1. Further, these two transient expression systems were used for the functional analysis of CtCHS1 as an example. After overexpression, relative CtCHS1 expression increased, particularly in Agrobacterium-transformed calli. Additionally, the contents of some flavonoids were altered; for instance, naringenin and genistein levels were significantly increased in Agrobacterium-transformed calli, whereas luteolin, luteolin-7-O-rutinoside, and apigenin derivative levels were significantly decreased in biolistic-transformed calli. Conclusion Using safflower callus as the experimental material, highly efficient Agrobacterium and biolistic transient expression systems were successfully established, and the utility of both systems for investigating gene function was demonstrated. The proposed safflower callus transient expression systems will be useful for further functional analyses of flavonoid biosynthetic genes in safflower.

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“…The expression of sweet potato CP2 altered the developmental and stress response characteristics of transgenic Arabidopsis plants [ 18 ]. However, Carthamus tinctorius CP1 acts as a negative regulator in the response of transgenic Arabidopsis to low-temperature stress; that is, inhibiting the expression of CtCP1 enhanced plant cold resistance [ 21 ]. Recently, we identified the existence of defense and stress response elements (e.g., TC-rich repeats) and MYB binding sites in maize PCP (designated as ZmCP03 ), implying a possible role in drought resistance [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

Modifying the Expression of Cysteine Protease Gene PCP Affects Pollen Development, Germination and Plant Drought Tolerance in Maize

Niu

Zhou

et al. 2023

IJMS

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Cysteine proteases (CPs) are vital proteolytic enzymes that play critical roles in various plant processes. However, the particular functions of CPs in maize remain largely unknown. We recently identified a pollen-specific CP (named PCP), which highly accumulated on the surface of maize pollen. Here, we reported that PCP played an important role in pollen germination and drought response in maize. Overexpression of PCP inhibited pollen germination, while mutation of PCP promoted pollen germination to some extent. Furthermore, we observed that germinal apertures of pollen grains in the PCP-overexpression transgenic lines were excessively covered, whereas this phenomenon was not observed in the wild type (WT), suggesting that PCP regulated pollen germination by affecting the germinal aperture structure. In addition, overexpression of PCP enhanced drought tolerance in maize plants, along with the increased activities of the antioxidant enzymes and the decreased numbers of the root cortical cells. Conversely, mutation of PCP significantly impaired drought tolerance. These results may aid in clarifying the precise functions of CPs in maize and contribute to the development of drought-tolerant maize materials.

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Identification and functional characterization of safflower cysteine protease 1 as negative regulator in response to low-temperature stress in transgenic Arabidopsis

Cited by 9 publications

References 58 publications

A Novel Senescence-Specific Gene (ZmSAG39) Negatively Regulates Darkness and Drought Responses in Maize

A Novel Senescence-Specific Gene (ZmSAG39) Negatively Regulates Darkness and Drought Responses in Maize

The establishment of transient expression systems and their application for gene function analysis of flavonoid biosynthesis in Carthamus tinctorius L

Modifying the Expression of Cysteine Protease Gene PCP Affects Pollen Development, Germination and Plant Drought Tolerance in Maize

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