scholarly journals Transcriptome Sequencing and Chemical Analysis Reveal the Formation Mechanism of White Florets in Carthamus tinctorius L.

Plants ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 847
Author(s):  
Tingyan Qiang ◽  
Jiushi Liu ◽  
Yuqing Dong ◽  
Yinbo Ma ◽  
Bengang Zhang ◽  
...  
Keyword(s):  
Transcriptome Sequencing ◽  
Phenylalanine Ammonia Lyase ◽  
Chalcone Synthase ◽  
Chemical Constituents ◽  
Carthamus Tinctorius ◽  
Cdna Libraries ◽  
Chain Reaction ◽  
Coa Ligase ◽  
Pigment Biosynthesis ◽  
Polymerase Chain

Carthamus tinctorius L. (safflower), an economic crop and herb, has been extensively studied for its diverse chemical constituents and pharmacological effects, but the mechanism of safflower pigments (SP) leading to different colors of florets has not been clarified. In the present study, we compared the contents of SP in two varieties of safflower with white and red florets, named Xinhonghua No. 7 (WXHH) and Yunhong No. 2 (RYH). The results showed the contents of SP in RYH were higher than WXHH. To investigate genes related to SP, we obtained six cDNA libraries of florets from the two varieties by transcriptome sequencing. A total of 225,008 unigenes were assembled and 40 unigenes related to safflower pigment biosynthesis were annotated, including 7 unigenes of phenylalanine ammonia-lyase (PAL), 20 unigenes of 4-coumarate-CoA ligase (4CL), 1 unigene of trans-cinnamate 4-monooxygenase (C4H), 7 unigenes of chalcone synthase (CHS), 4 unigenes of chalcone isomerase (CHI), and 1 unigene of flavanone 3-hydroxylase (F3H). Based on expression levels we selected 16 differentially expressed unigenes (DEGs) and tested them using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR), which was consistent with the sequencing results. Consequently, we speculated that in WXHH, 3 PALs, 3 4CLs, 1 C4H, 1 CHS, and 1 CHI, which were down-regulated, and 1 F3H, which was up-regulated, may play a key role in the formation of white florets.

scholarly journals Differential Expression of Benzophenone Synthase and Chalcone Synthase in Hypericum Sampsonii

2012 ◽  
Vol 7 (12) ◽  
pp. 1934578X1200701 ◽  
Author(s):  
Lili Huang ◽  
Hong Wang ◽  
Hechun Ye ◽  
Zhigao Du ◽  
Yansheng Zhang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Differential Expression ◽  
Chalcone Synthase ◽  
Quantitative Polymerase Chain Reaction ◽  
Transcript Level ◽  
Reproductive Stage ◽  
Vegetative Stage ◽  
Chain Reaction ◽  
Expression Levels ◽  
Polymerase Chain

cDNAs encoding Hypericum sampsonii benzophenone synthase (HsBPS) and chalcone synthase (HsCHS) were isolated and functionally characterized. Differential expressions of HsBPS and HsCHS were monitored using quantitative polymerase chain reaction (PCR). In the vegetative stage, HsBPS was highly expressed in the roots; its transcript level was approx. 100 times higher than that of HsCHS. Relatively high transcript amounts of HsBPS were also detected in older leaves, whereas the youngest leaves contained higher transcript amounts of HsCHS. In the reproductive stage, maximum HsCHS expression was detected in flowers, the transcript level being approx. 5 times higher than that of HsBPS. The inversed situation with a 10-fold difference in the expression levels was observed with fruits. High transcript amounts for both proteins were found in roots.

scholarly journals 560 Molecular Analysis of the First Intron of the Chalcone Synthase A Gene in Petunia

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 542F-543
Author(s):  
R.J. Griesbach ◽  
R. Beck
Keyword(s):  
Genetic Diversity ◽  
Chalcone Synthase ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Pcr Primers ◽  
Chain Reaction ◽  
First Intron ◽  
Single Fragment ◽  
Polymerase Chain ◽  
Restriction Fragment Length

A new method was developed to analyze genetic diversity among Petunia species. The first intron of the chalcone synthase A gene was cloned through the polymerase chain reaction (PCR) and partially sequenced. This sequence was used to dissect the intron into two halves (3' and 5' halves). The PCR primers for the 5' half amplified a single fragment that was the same length for all of the species that were studied. Restriction fragment length polymorphism (RFLP) analysis of the 5' half resulted in the same number and length of fragments for all the species that were evaluated. The PCR primers for the 3' half amplified a number of fragments that were characteristic for each species. This research provides a new tool for measuring genetic diversity. Genetic diversity measured with this tool should be closely related to evolutionary distance.

scholarly journals Differential inductions of phenylalanine ammonia-lyase and chalcone synthase during wounding, salicylic acid treatment, and salinity stress in safflower, Carthamus tinctorius

2014 ◽  
Vol 34 (3) ◽  
Author(s):  
Sara Dehghan ◽  
Mahnaz Sadeghi ◽  
Anne Pöppel ◽  
Rainer Fischer ◽  
Reinhard Lakes-Harlan ◽  
...  
Keyword(s):  
Salicylic Acid ◽  
Salinity Stress ◽  
Phenylalanine Ammonia Lyase ◽  
Chalcone Synthase ◽  
Expression Profiles ◽  
Carthamus Tinctorius ◽  
Rt Pcr ◽  
Defence Mechanisms ◽  
Similar Expression Pattern ◽  
Defence Signalling

Safflower (Carthamus tinctorius L.) serves as a reference dicot for investigation of defence mechanisms in Asteraceae due to abundant secondary metabolites and high resistance/tolerance to environmental stresses. In plants, phenylpropanoid and flavonoid pathways are considered as two central defence signalling cascades in stress conditions. Here, we describe the isolation of two major genes in these pathways, CtPAL (phenylalanine ammonia-lyase) and CtCHS (chalcone synthase) in safflower along with monitoring their expression profiles in different stress circumstances. The aa (amino acid) sequence of isolated region of CtPAL possesses the maximum identity up to 96% to its orthologue in Cynara scolymus, while that of CtCHS retains the highest identity to its orthologue in Callistephus chinensis up to 96%. Experiments for gene expression profiling of CtPAL and CtCHS were performed after the treatment of seedlings with 0.1 and 1 mM SA (salicylic acid), wounding and salinity stress. The results of semi-quantitative RT–PCR revealed that both CtPAL and CtCHS genes are further responsive to higher concentration of SA with dissimilar patterns. Regarding wounding stress, CtPAL gets slightly induced upon injury at 3 hat (hours after treatment) (hat), whereas CtCHS gets greatly induced at 3 hat and levels off gradually afterward. Upon salinity stress, CtPAL displays a similar expression pattern by getting slightly induced at 3 hat, but CtCHS exhibits a biphasic expression profile with two prominent peaks at 3 and 24 hat. These results substantiate the involvement of phenylpropanoid and particularly flavonoid pathways in safflower during wounding and especially salinity stress.

PCR-generated cDNA libraries from reduced numbers of mouse oocytes

Zygote ◽  
1995 ◽  
Vol 3 (3) ◽  
pp. 241-250 ◽  
Author(s):  
Frédérique Revel ◽  
Jean-Paul Renard ◽  
Véronique Duranthon
Keyword(s):  
Genetic Information ◽  
Preimplantation Embryos ◽  
Cdna Libraries ◽  
Cdna Synthesis ◽  
Chain Reaction ◽  
Mouse Oocytes ◽  
Cdna Population ◽  
Cellular Extract ◽  
Reproducible Method ◽  
Polymerase Chain

SummaryWe describe a rapid and reproducible method for cloning cDNA amplified from 10 mouse oocytes. The procedure consists in priming cDNA synthesis from a crude cellular extract using an oligo d(T) containing primer and submitting the size-limited cDNA first strand to poly(dG) tailing. The whole cDNA population is then polymerase chain reaction (PCR) amplified using two primers complementary to oligo d(A) and oligo d(G) ends of the cDNA. In this procedure no purification steps are required. We obtained about 5 ×106 clones from 10 oocytes. Screening of the library showed that the relative abundance of the transcripts was preserved during amplification and cloning and that the procedure allows cloning of low-abundance sequences at least as rare as 0.008% of the mRNA. The repeatable generation of representative cDNA libraries from reduced numbers of oocytes or embryos should open new opportunities for obtaining genetic information from mammalian preimplantation embryos.

scholarly journals Molecular Heterogeneity of the Chalcone Synthase Intron in Petunia

HortScience ◽  
2000 ◽  
Vol 35 (7) ◽  
pp. 1347-1349 ◽  
Author(s):  
R.J. Griesbach ◽  
R.M. Beck ◽  
J.R. Stehmann
Keyword(s):  
Species Complex ◽  
Chalcone Synthase ◽  
Molecular Heterogeneity ◽  
Chain Reaction ◽  
Chalcone Synthase Gene ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Species Specific ◽  
Degree Of Heterogeneity ◽  
Petunia Integrifolia

A method was developed to characterize the genetic heterogeneity of the chalcone synthase gene intron within the Petunia integrifolia (Hook.) Schinz & Thell. species complex. The DNA from wild species collected from known locations was used to amplify the chalcone synthase gene intron through the polymerase chain reaction (PCR). The resulting PCR product was then characterized by Rsa 1 restriction, revealing a degree of heterogeneity that could be used to characterize the species genetically. Of the four different species that were characterized, two could be placed in the same genetic grouping. This study shows that the variation in the intron of the Chs A gene may be species-specific.

scholarly journals Rapid Freeze Acclimation of Poncirus trifoliata Seedlings Exposed to 10 °C and Long Days

HortScience ◽  
1997 ◽  
Vol 32 (5) ◽  
pp. 854-857 ◽  
Author(s):  
Milton E. Tignor ◽  
Frederick S. Davies ◽  
Wayne B. Sherman ◽  
John M. Davis
Keyword(s):  
Water Potential ◽  
Messenger Rna ◽  
Phenylalanine Ammonia Lyase ◽  
Chalcone Synthase ◽  
Poncirus Trifoliata ◽  
Time Interval ◽  
Tissue Samples ◽  
Complementary Dna ◽  
Coa Ligase ◽  
Red Coloration

Poncirus trifoliata (L.) Raf. seeds were germinated in perlite under intermittent mist at about 25 °C and natural daylight in a greenhouse. Two-week-old seedlings were then transferred into a growth chamber at 25 °C and 16-hour daylength for 1 week. Tissue samples were collected at 0, 6, 24, 168, and 504 hours after temperature equilibration at 10 °C. Freezing tolerance at –6.7 °C, as determined by electrolyte leakage, and stem (leaves attached) water potential (ψx), measured using a pressure chamber, was recorded for a subset of seedlings for each time interval. Red coloration (apparently anthocyanin) developed at the petiole leaflet junction and buds after 48 hours at 10 °C and gradually occurred throughout the leaves during further exposure. Complementary DNA clones for phenylalanine ammonia lyase (PAL), 4-coumarate: coA ligase (4CL), and chalcone synthase (CHS) were used to probe RNA isolated from the leaves. No increase in steady-state messenger RNA level was detected. Increases in freeze hardiness occurred within 6 hours in the leaves, and continued for up to 1 week. Water potential initially decreased from –0.6 to –2.0 MPa after 6 hours, then returned to –0.6 MPa after 1 week. Thus, Poncirus trifoliata seedlings freeze-acclimate significantly after only 6 hours at 10 °C.

Polymerase chain reaction-based construction of cDNA libraries from minute amounts of third-stage and fourth- and fifth-stage larvae of Dictyocaulus viviparus

1996 ◽  
Vol 83 (1) ◽  
pp. 20-23 ◽  
Author(s):  
G. von Samson-Himmelstjerna ◽  
Gero Wunderlich ◽  
F. Mühlschlegel ◽  
Matthias Frosch ◽  
Thomas Schnieder
Keyword(s):  
Polymerase Chain Reaction ◽  
Cdna Libraries ◽  
Chain Reaction ◽  
Dictyocaulus Viviparus ◽  
Third Stage ◽  
Polymerase Chain
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