Breeding Of New Strains Of Colored Potato And HPLC-MS Analysis Of Anthocyanin Components

Colored potato is rich in anthocyanins and other nutrients,which is not only a characteristic crop,but also an important raw material for extracting natural pigment.Anthocyanins have strong free radical scavenging and antioxidant function,with great value in human health,medicine,food and additive of cosmetics.At present,there is severe lack of superior colored potato varieties with high yield,high anthocyanin content,prominent comprehensive character,and the potential to promote and grow in large areas at home and abroad.In recent years,in order to breed new colored potato varieties with high anthocyanin content and prominent agronomic characters,according to the matching principle of large difference of ecotype,trait complementary between parents,we used the tetraploid varieties MIN-021,Red-P1 and Russet-01 introduced from America crossed with the domestic variety Heimeiren,obtained hybrid F1 seeds of "MIN-021×Heimeiren","Red-P1×Heimeiren" and "Russet-01×Heimeiren" by emasculation,bagging and and artificial pollination.After greenhouse cultivation and field selection,we selected five new colored potato hybrid strains Mengcai-01,Mengcai-02,Mengcai-03,Mengcai-04 and Mengcai-05,which had good comprehensive performance.In this research,using the red colored potatoes Hongmei and CY-1 as the control,we focused on the analysis and evaluation of yield,nutritional quality characteristics,drought tolerance,pollen fertility,chromosome pairing configuration,SSR fingerprint feature and genetic diversity of the five new strains.And identified the anthocyanin components and contents of Mengcai-01(with the relatively high anthocyanin content)and Mengcai-02(with the relatively low anthocyanin content)by the method of high performance liquid chromatography and mass spectrometry(HPLC-MS).The main results were as follow:1.Five new colored potato strains were selected from F]segregation individuals of the three hybrid combinations,named Mengcai-01,Mengcai-02,Mengcai-03,Mengcai-04 and Mengcai-05,of which the plant types were erect,the tubers were concentrated and neat with shallow eyes,elliptic or oblong shape,and the tuber flesh showed different degrees of purple.2.The new strain Mengcai-04 is medium early mature type,Mengcai-01 is medium late mature type,and Mengcai-02,Mengcai-03 and Mengcai-05 are medium mature type.All the five new strains have high average yield per plant and commodity rate of potato,with more than 1.11 kg and 80.66%,respectively.3.The anthocyanin content of new strain Mengcai-01 was up to 626.71 mg/kg·FW,Mengcai-02,Mengcai-03 and Mengcai-05 were more than 200 mg/kg·FW,and Mengcai-04 was 118.15 mg/kg-FW.The new strains Mengcai-01 and Mengcai-02 had a relatively high tuber dry matter content(21.32%and 22.38%)and starch content(17.11%and 18.81%),and other three new strains had more than 19%of dry matter content and 15%of starch content,respectively.The crude protein content of the five new strains ranged from 0.98%to 1.67%,the content of reducing sugar ranged from 0.015%to 0.028%,the content of Vc ranged from 18.67%to 21.20%,and the content of phenolic compounds ranged from 0.12%to 0.19%.4.The pollen fertility of Mengcai-01,Mengcai-02 and Mengcai-05 were more than 76.07%,and Mengcai-03 and Mengcai-04 were 33.39%and 58.74%,respectively.All the five new coloered potato strains can be used as the intermediate material for hybrid breeding.The chromosome configuration of new strain Mengcai-01 was 1.18 Ⅰ+14.65 Ⅱ+1.28Ⅲ+3.42Ⅳ,Mengcai-02 was 1.33 Ⅰ+13.97Ⅱ+1.22Ⅲ+3.77Ⅳ,Mengcai-03 was 9.12 Ⅰ+7.71 Ⅱ+6.88Ⅲ+0.7Ⅳ,Mengcai-04 was 3.96 Ⅰ+11.25 Ⅱ+4.91Ⅲ+1.7Ⅳ,and Mengcai-05 was 3.82 Ⅰ+13.05 Ⅱ+2.31Ⅲ+2.79Ⅳ.5.A total of 328 polymorphic loci were amplified by 31 screened suitable SSR primers,accounted for 89.86%of all the loci.The DNA fingerprints of five new strains,their parents and the control varieties were constructed by SSR primers C54,C57,STI027 and S153.Based on the GD value of 0.40,ten potato materials were classified into three categories:the new strains Mengcai-05,Red-P1,MIN-021 and CY-1 as a category;Mengcai-01 and Mengcai-02,Mengcai-03,Mengcai-04 and Heimeiren as a category;and Hongmei as a single category.6.The regeneration systems of shoot tip culture virus-free of the new potato strains were established,and the screened medium with optimum matching of hormone concentration was MS+6-BA0.1 mg/L+NAA0.2 mg/L+GA30.1 mg/L.The virus-free plantlets of five new strains were obtained according to DAS-ELISA detection,which laid the foundation for the propagation of virus-free seedlings and seeds.7.Using different concentrations of PEG solution as the treatment for new strains of virus-free tissue culture seedlings of Mengcai-01,Mengcai-02 and Mengcai-05 and their parents,the relative membrane permeability,MDA content and proline content of seedling leaves were determined under different drought stress intensity.After comprehensive evaluation,the orders of drought resistance ability of the six materials were Mengcai-01>Mengcai-05>MIN-021>Red-P1>Mengcai-02>Heimeiren.8.The results of HPLC-MS analysis showed that new strains Mengcai-01 and Mengcai-02 with purple flesh contain petunidin-3-(4’"-p-coumarylrutinoside),petunidi n-3-(4"’-p-coumarylrutinoside)-5-glucoside,petunidin-3-(4"’-feruloylrutinoside)-5-glucosi de,of which petunidin-3-(4"’-p-coumarylrutinoside)-5-glucoside was the main compon ent.In addition,Mengcai-01 was unique to malvidin-3-(4"’-p-coumarylrutinoside-5-gl ucoside),and Mengcai-02 was unique to peonidin-3-(4’"-p-caffeylrutinoside)-5-glucosi de and peonidin-3-(4"’-p-coumarylrutinoside)-5-glucoside.The control variety Hongme i with red flesh contains pelargonidin-3-rutinoside,cyanidin-3-coumaryl-glucoside,pel argonidin-3-(4"’-feruloylrutinoside)-5-glucoside and pelargonidin-3-(4"’-p-coumarylrutino side)-5-glucoside,of which pelargonidin-3-(4"’-p-coumarylrutinoside)-5-glucoside and cyanidin-3-coumary-glucoside were the main components.