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Journal of Integrative Agriculture
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Functional analysis of the nitrogen metabolism-related gene
CsGS1
in cucumber
XIN Ming, QIN Zhi-wei, YANG Jing, ZHOU Xiu-yan, WANG Lei
2021, 20 (
6
): 1515-1524. DOI:
10.1016/S2095-3119(20)63305-6
Abstract
(
128
)
PDF in ScienceDirect
Glutamine synthetase (GS) plays an important role in nitrogen (N) metabolism in cucumber. In this study, we cloned and sequenced the CsGS1 gene, and analyzed the expression patterns and subcellular localization of the GS1 protein in response to different N conditions in order to determine its role in low-nitrogen (LN) tolerance.
CsGS1
was abundantly expressed in the leaves of the low N-requiring cultivar D0328, while the high N-requiring cultivar D0422 showed similar expression levels across different tissues including leaves, shoots and roots. Furthermore, the GS1 protein was primarily localized in the cytoplasm of plant cells. Both cultivars were then transformed with the
CsGS1
coding sequence or antisense sequence
via
Agrobacterium
tumefaciens
in order to overexpress and silence
GS1
expression, respectively. Overexpression of
CsGS1
significantly improved LN tolerance and photosynthetic parameters, and increased chlorophyll b content, biomass, plant height, root length, N accumulation and GS activity under LN condition compared to the control.
CsGS1
silencing on the other hand significantly reduced the above indices. Taken together,
CsGS1
is crucial for maintaining N metabolism in cucumber plants during N deprivation, and is a promising target for generating novel transgenic breeds with increasing nitrogen utilization efficiency.
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Proteomic Analysis of Fruit Bending in Cucumber (Cucumis sativus L.)
WANG Li-li, ZHANG Peng, QIN Zhi-wei , ZHOU Xiu-yan
2014, 13 (
5
): 963-974. DOI:
10.1016/S2095-3119(13)60406-2
Abstract
(
1934
)
PDF in ScienceDirect
In cucumber, fruit shape is an important quality criterion, and fruit bending is known to limit growth, yield, and taste. To investigate the post-transcriptional changes that regulate fruit bending and to better understand the underlying molecular mechanisms, we generated a proteomic profile of the abdomen and back of cucumber bending fruit. Two-dimensional gel electrophoresis (2-DE) allowed the detection of approximately 900 distinct protein spots in each gel, 32 of which were differentially expressed in the abdomen and back of bending cucumber fruit. Ten of the differentially expressed proteins were analyzed using matrix-assisted laser ionization time of flight mass spectrometry (MALDI-TOF/MS). A search of primary databases showed that the identified proteins are involved in various metabolic processes and cellular responses, including photosynthesis metabolism, energy metabolism, defense and stress response, and regulation. The identified proteins included large subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase, which are involved in photosynthesis and photorespiratory metabolism, and isocitrate dehydrogenase, which is involved in the tricarboxylic acid cycle. It is possible that imbalances in catabolic and anabolic processes directly affect the bending of cucumber fruit. The predicted function of the cobalamin- independent methionine synthase isozyme is closely related to ethylene biosynthesis; fruit bending may be regulated by ethylene, or by ethylene signaling crosstalk during fruit development. The 14-3-3 protein is usually considered to be a regulation-related protein, which plays a role in regulating cell hyperplasia, cell differentiation during growth, and apoptosis during senescence. Involvement of guanosine triphosphate (GTP)-binding proteins in signal transmission is known to regulate the development of cells in cucumber fruits and to play a role in fruit shape variation. Patterns of protein expression showed high repeatability. We hypothesize that these proteins may play an important role in growth and bending of cucumber fruits. The results of our study provide insight into the genetic mechanism underlying fruit bending in cucumber, and may help to promote cultivation of new varieties with superior fruits.
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Proteomic Analysis of Cucumber Defense Rresponses Induced by Propamocarb
WU Peng, QIN Zhi-wei, WU Tao, ZHOU Xiu-yan, XIN Ming , GUO Qian-qian
2013, 12 (
11
): 2022-2035. DOI:
10.1016/S2095-3119(13)60370-6
Abstract
(
1414
)
PDF in ScienceDirect
Propamocarb is an agricultural chemical that has been widely used to protect cucumber plants from downy mildew. To understand the mechanisms of cucumber defense responses to propamocarb, we investigated the physiological and proteomic responses of the cucumber line D0351 with propamocarb application. We found that after treatment with propamocarb, the activities of detoxifying enzymes (glutathione reductase, GR; glutathione S-tramsferase, GST) and soluble sugar content of cucumber fruit were significantly increased, but malonaldehyde (MDA) content was significantly reduced. To identify components of propamocarb responsive signaling, we compared the high resolution two-dimensional gel electrophoresis (2-DE) protein profiles of control and propamocarb-treated fruits, and identified 18 differentially expressed (13 up-regulated and 5 down-regulated) proteins induced by propamocarb which were determined by matrix-assisted laser desorption/ionization timeof- flight mass spectrometry (MALDI-TOF-MS). The majority of the proteins had functions related to detoxication, energy and transport, protein biosynthesis, regulating reactions and defending against stresses. A real-time quantitative reverse transcriptional-polymerase chain reaction (qRT-PCR) was used to compare transcript and protein accumulation patterns for 18 candidate proteins, and the expression of 14 was consistent at both transcript and protein levels. The responses of cucumber proteome to propamocarb seemed complex; the identified proteins may play an important role in regulating adaptation activities following exposure to propamocarb. Data presented herein may shed light on understanding cucumber fruit defense responses under propamocarb treatment.
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