| Purpose:Panax notoginseng as rare Chinese medicine.Because of the influence of continuous cropping obstacles and market requirement,now P.notoginseng cultivation presents a trend of introduction to the high-altitude and low-temperature regions.In order to provide theoretical basis for the introduction and planting of P.notoginseng,the experiment was conducted at different temperatures on the total saponins and photosynthetic physiology of P.notoginseng.And also studied the effects of different temperatures on the sequencing results of 3-year old P.notoginseng genes.Methods:1.This study analyzed the changes of saponins content of 1,2,3 years old P.notoginseng root in 30 days treatment under 10?,15? and 20?.2.1,2 and 3 years old P.notoginseng were treated at 10?,15? and 20? for 30 days.The photosynthetic parameters were determined by LI-COR 6400 portable photosynthetic analyzer.MDA was determined by TAB colorimetric method and the photosynthetic pigment content was determined at 3 temperatures.3.The 3-year P.notoginseng was treated at 10?,15? and 20? for 30 days,and the3-year P.notoginseng with normal growth and consistent growth in the planting base(CK as the control group).HPLC was used to determine the contents of 4 monomer saponins and their total saponins in the four parts of its main root,rhizome,stem and leaf.4.The 3-year P.noginseng treated at 10?,15? and 20? for 30 days,and the 3-year P.noginseng treated at 10?,15? and 20? with normal growth and consistent growth in the planting base(CK as the control group)were sequenced on Illumina NovaSeq 6000 platform and PacBio Sequel platform.Results:1.By comparing the contents of total saponins in roots of P.notoginseng aged 1 year,2 years and 3 years for 30 days at 10?,15? and 20?,it was found that 10?,15? had no significant effect on the accumulation of saponins in roots of P.notoginseng aged 2 and3 years,and the optimal temperature for the accumulation of P.notoginseng was 20?.The content of P.notoginseng was the highest at 10?,and the low temperature was conducive to the accumulation of P.notoginseng.2.By comparing the photosynthetic physiology of P.notoginseng grown in 1,2 and 3years under 10?,15? and 20? for 30 days,the photosynthetic physiology of P.notoginseng decreased with the decrease of temperature.2 and 3-year P.notoginseng had the highest MDA content at 10?,and 1-year P.notoginseng had the highest MDA content at 15?.2 years of notoginseng Chla,Chlb and carotenoid were highest in 15 ?;The photosynthetic physiology of P.notoginseng in different years was different at low temperature.3.HPLC method was used to compare the contents of saponins in the 3 years P.notoginseng cultured for 30 days at 10?,15? and 20?,as well as in the CK group.It was found that the content of total saponins in P.notoginseng was the highest at 20?,and the content of total saponins in P.notoginseng at 15? and 10? was also higher than that in CK group.The content of total saponins in rhizome was the highest at 15?,and the content of total saponins in rhizome was also higher than that in CK group at 20? and10?.The content of total saponins in P.notoginseng leaves was the highest at 15? and the lowest at 20?.At the same time,the total content of notoginseng saponins in the harvest period was higher than that in the CK group harvested in advance.4.Two-generation sequencing and three-generation sequencing of P.notoginseng leaves were used to read 729094 polymerases on PacBio Sequel platform,with a total data of 25.80 GB.Moreover,Isoform annotated to 34990 in NR,NT,GO,KOG,Pfam,KEGG and Swissprot seven functional databases.It successfully annotated to 21959 novel genes in GO,KEGG,NR,Swissprot,Trembl and eggNOG.Among the second-generation sequencing results,the total variable shear event was the highest in the 15? group.Amongthe sequencing results of the third generation,the variable shear form accounted for 60.4%of the transcript.At the same time,the gene MSTRG.10001 was only compared in the 10?group,and the other three groups were not matched.Through NT and NR annotation,it was found that P.notoginseng gene had similar ratio with daucus carota,vitis vinifera,sesamum indicum,coffea canephora,etc.KOG found that general function predicted only is the most genes.GO classification showed that the most differentially expressed genes were found at 20? vs 10?.KEGG found that the leaf of P.notoginseng had the most genes involved in metabolism.The gene specific to NR was found to be the most by SwissPort annotation.Conclusion:Effects of temperature on saponins accumulation of P.notoginseng with different growth years were different.Among them,15??10? are conducive to the accumulation of saponins in 1-year P.notoginseng and the second-generation sequencing results show that the 15? group has the highest total variable shear event.2,3 years old P.notoginseng,at 20? it was beneficial to its saponins accumulation.Hypothermia is not conducive to photosynthesis of P.notoginseng,and low temperature will cause the content increase of MDA in P.notoginseng.In three-year-old P.notoginseng,the content of saponins in the main roots and leaves of P.notoginseng at low temperature was different,but the rhizome and stems were not sensitive to temperature.Through the second and third generation of P.notoginseng leaf sequencing,it is beneficial to further study the effect of temperature on the synthesis of notoginseng saponin related genes.And lays a foundation for the next P.notoginseng genome research. |
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