Background The Agrobacterium -mediated transient transformation is a versatile and indispensable way of rapid analyzing gene function in plants. Despite this transient expression system has been successfully applied in a number of plant species, it is poorly developed in Caragana intermedia . Results In this study, we established an Agrobacterium -mediated transient expression system in C. intermedia leaves and optimized the effect of different Agrobacterial strains, several surfactants and the concentration of Silwet L-77, which would affect transient expression efficiency. Among the 5 Agrobacterial strains examined, GV3101 produced the highest GUS expression level. Besides, higher level of transient expression was observed in plants infiltrated with Silwet L-77 than with Triton X-100 or Tween-20. Silwet L-77 at a concentration of 0.001% greatly improved the level of GUS transient expression. Real-time PCR showed that expression of CiDREB1C was highly up-regulated in transiently expressed plants and reached the highest level at the 2nd day after infiltration. Based on this optimized transient transformation method, we characterized CiDREB1C function in response to drought, salt and ABA treatment. The results showed that transiently expressed CiDREB1C in C. intermedia leaves could enhance the survival rate and chlorophyll content, and reduce the lodging rate compared with the control seedlings under drought, salt and ABA treatments. Furthermore, the rate of leaf shedding of CiDREB1C transient expression seedlings was lower than that of the control under ABA treatment. Conclusions The optimized transient expression condition in C. intermedia leaves were infiltrated with Agrobacterial strains GV3101 plus Silwet L-77 at a concentration of 0.001% added into the infiltration medium. Transiently expressed CiDREB1C enhanced drought, salt and ABA stress tolerance, indicated that it was a suitable and effective tool to determine gene function involved in abiotic stress response in C. intermedia . Electronic supplementary material The online version of this article (10.1186/s12870-019-1800-4) contains supplementary material, which is available to authorized users.
Caragana intermedia, a leguminous shrub widely distributed in cold and arid regions, is rich in secondary metabolites and natural active substances, with high nutritional and medical values. It is interesting that the pods of C. intermedia often show different colors among individual plants. In this study, 10-, 20- and 30-day-old red and green pods of C. intermedia were used to identify and characterize important metabolites associated with pod color. A total 557 metabolites, which could be classified into 21 groups, were detected in the pod extracts using liquid chromatography coupled with ESI-triple quadrupole-linear ion trap mass spectrometer (LC-ESI-MS/MS). Metabolomics analysis revealed significant differences in 15 groups of metabolites between red and green pods, including amino acids, nucleotide derivatives, flavonoids, and phytohormones. Metabolic pathway analysis showed that the shikimic acid and the phytohormone metabolic pathways were extraordinarily active in red pods, and the difference between red and green pods was obvious. Moreover, red pods showed remarkable flavonoids, cytokinins, and auxin accumulation, and the content of total flavonoids and proanthocyanidins in 30-day-old red pods was significantly higher than that in green pods. This metabolic profile contributes to valuable insights into the metabolic regulation mechanism in different color pods.
Caragana. intermedia Kuang et H.C.Fu., a xerophytic deciduous shrub that widely distributed in arid and semi-arid desert areas of North-west China, is highly tolerant to cold, drought, salt-alkali and barren. In this study, the transcriptome C. intermedia under drought treatment was screened for the genes encoding the AP2/ERF family of transcription factors. As a result, 22 sequences with complete open reading frames (ORFs) were obtained. All sequences were divided into 13 groups, including the DREB A1 to A6, ERF B1 to B6, and RAV groups. The results of multiple sequence alignment (MSA) analysis, domain distribution analysis, conservative motif analysis and tertiary structure prediction showed that all the AP2/ERF transcription factors contained the AP2 domain. The AP2 domain contained the YRG and RAYD elements. In addition, quantitative real-time polymerase chain reaction (qRT-PCR) was employed to analyze the expression patterns of the 22 AP2/ERF transcription factors in C. intermedia. Most of the transcription factors exhibited tissue-speci c expression. The expression of most transcription factors was induced to varying degrees by cold, heat, salt and mannitol stress. Furthermore, Heterologous expression of CiDREB3 in transgenic Arabidopsis L. Heynh decreased root length and fresh weight, decreased rosette leaf diameter and number of leaf, and improved drought tolerance during seedlings development. Taken together, the results of the present study help to better understand the functions of the AP2/ERF family of transcription factors in plant responses to multiple abiotic stresses and deeply insight the roles of CiDREB3 in Arabidopsis morphology and conferring abiotic stress tolerance.
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