Optimization Of Key Factors In The Measurement Of Quality Of Porphyra Haitanensis And Quality Analysis Of Its New Lines

This article studied the test conditions of Porphyra Haitanensis major characters, includingphycobiliprotein, chlorophyll, crude protein, total amino acid and free amino acid, and conductedan optimization on it. The results gained a series of scientific accurate empirical method ofPorphyra Haitanensis quality. And carried a characters anaylsis on different filial generations ofsix new lines that came from hybridization and selection and control line by this way, they werehigh-production line(Z-17), good quality line(Q-1), high temperature resistance line(Z-26), thinthallus line(Z-61), low nitrogen(N) an phosphorus(P) resistance line(9-Ⅳ) and low salinityresistance line(8-Ⅰ) respectively. It layed foundation for germplasm improvement and selectionof the high-quality new lines through character analysis. Furthermore, this paper took analysis oncomposition of protein in these new lines by SDS electrophoretic technique, then provided a newway to improve and identify germplasm on the level of protein.1、The optimization of the new lines test conditions: used Orthogonal experimental design tooptimize and the results as follows: It combinated trituration and freeze thawing repeatly todestroy cell wall and extract phycobiliprotein, extractant was0.05mol/L phosphate buffer saline,thallus quality was0.010g and it was supposed to avoid illumination during extraction, thenmensurate content after six time freeze thawing. It applied trituation to extract chlorophyll A,thallus quality was also0.010g and it must avoid illumination too during extraction. It was bestto weigh0.150g thallus to extract crude protein with extractant10ml sulpuric acid. Using0.050gthallus to extract total amino acid, the extractant was0.1%hydrochloric acid, after drying under60℃, then diluting three times and anlysis automaticly by instrument. Similarly, using0.010gthallus to extract free amino acid, the extractant was deionized water, the time of extraction was4h, the constant temperature was60℃.2、Qualitative characters of quick-grow line(Z-17): There was quiet difference among thephycobiliprotein level of its four filial generations thalli, but they all significant higher than thecontrol(68.01±3.33)mg/g. The fourth filial generation(F4) possessed highestlevel(123.35±3.69)mg/g, which was81%higher than the control; The contents of the chlorophyllA in these four generations thalli were obviously high than the control, and in F2thalli was thehighest, which up to (9.78±0.03)mg/g, was48.18%higher than the latter(6.60±0.36)mg/g. Crudeprotein level was likewise quite different from the control, and in F2thalli was the highest, whichwas35.71%higher than the control, up to (444.95±6.44)mg/g; The total amino acid levels of the four generations that have an aboundant content of essential amino acid for human were alldistinctly higher than the control, especially the content of Val, Leu, Lys and so on; Except Ala,in F4thalli the its level was10.93mg/g, which was1.24times higher than the control, all of otherfree amino acid levels were quiet different from the latter.3、Qualitative characters of good quality line(Q-1): The contents of the total phycobiliproteinand chlorophyll A levels in its four filial generations were both significant higher than the control,and in F4thalli were the highest, which up to (120.06±1.84)mg/g and (10.39±0.08)mg/grespectively. Either in the experimental or the control thalli, the levels of the phycobiliproteinand chlorophyll A showed positive correlation. The content of crude protein, total amino acid,and free amino acid were all sharply higher than the control. For example, the crude protein levelin F4was high to (444.72±5.30)mg/g, which was35.6%obviously higher than the control; thetotal amino acid level was (385.31±5.22)mg/g, which was33.5%obviously higher than thecontrol; Moreover, the contents of the four tastable amino acid(Asp, Glu, Ala, Gly) weremarkedlly higher than the control.4、Qualitative characters of new thin thalli line (Z-61): The phycobiliprotein and chlorophyllA contents in four filials were obviously higher than the control without harvesting and at thefirst harvesting; The content variation of crude protein in the same filial was: the firstharvesting>without harvesting>the second harvesting, and there was no obvious variationbetween the first harvesting and without harvesting. The crude protein content of the thalli in allfilials at first harvesting and without harvesting were obviously higher than the control. The totalamino acid and free amino acid contents at the first harvesting and without harvesting wereobviously higher than the second harvesting in four filials, and the flavor amino acids contentswere abundant.5、 Qualitative characters of new high temperature resistance line (Z-26): Thephycobiliprotein and chlorophyll contents in all filials were obviously higher than the control,and the highest phycobiliprotein content was (142.29±2.75)mg/g in F4which was2.09timesthan the control. Also the chlorophyll A content reached to (10.51±0.89)mg/g. The crude proteincontent was higher or obviously higher than the control. The total amino acid content wasobviously higher than the control. And it was rich in essential amino acid, especially Val., Leu.and Lys., which were higher than the control. The total content of free amino acid was obviouslyhigher than the control. the flavor amino acids including Asp., Glu. and Phe. are rich. In hightemperature resistance experiment, all the index were fluctuated at26℃and21℃for10daysand descended at30℃.6、Qualitative characters of new low N&P resistance line (9#IV): Five qualitative characters(phycobiliprotein, chlorophyll A, crude protein, total amino acid, free amino acid) were descending with increasing cultivated days under low N&P condition and there was obviousvariation in quality index between the thalli cultivated under low N&P condition or not. Fivequalitative characters of the thalli that cultivated under low N&P condition for ten daysdescended to32.5%、24.8%、37.7%、40.8%、26.3%of the normal thalli.7、Qualitative characters of new low salinity resistance line (8#I): The phycobiliproteincontent of F3thalli was higher than F1or F2. And the highest content was (86.41±0.60)mg/gunder normal salinity, which was a little more higher than the content under low salinity. Thehighest chlorophyll A content was (7.52±0.24)mg/g, the thalli of which were cultivated underlow sanility(10). The crude protein contents of F1and F3under normal salinity were a littlehigher than the ones under low salinity condition. And the highest crude protein content of F3was (390.63±8.53)mg/g under normal salinity. The highest total amino acid content of F3was(317.88±16.74)mg/g under normal salinity. The more lower salinity, the less content of totalamino acid. The free amino acid content of the thalli reached to (24.90±0.57)mg/g which waslittle higher than the control under normal salinity.8、SDS electrophoresis of Porphyra Haitanensis: Six common bands were separated fromseven samples of the Porphyra Haitanensis and the location of16-20KDa was a thick band. Theother five bands were55.0、48.0、42.0、38.0、30.0KDa. There was a special band(a) at12.0KDaof sample one. There was a special band(b) at28.0KDa of sample two. There was no obviousband at20.0KDa of sample five.