Effect of Mineral Elements on Phenotypic Diveristy, Fruit-body Yield, Anamorph Diveristy and Activity of Related Enzymes of Colony Types of Cordyceps Militaris in Subculturing
|School||Shenyang Agricultural University|
|Keywords||Cordyceps Mineral elements Colonies corners Fruiting bodies Strain degradation Subculture|
In this study, using orthogonal test, the North Cordyceps excellent strains L20 in different mineral elements medium subcultured 12 generations, seven different types of colonies, colony morphology of the different types of diversity, maintaining algebra, mycelial growth rate dry weight of mycelium, fruiting body production, the angle variable rate, clonal diversity, sporulation, esterase activity, SOD activity and cellulase activity of research, to explore the mineral elements subculture colony phenotype changes in vegetative diversity, and enzyme activity and fruiting body production. The main results are as follows. Strain L20 by multiple adapter medium of different mineral elements can occur the apparent angular phenomena, the formation of seven different types of colonies, fruiting bodies of colony forming ability of I, II, Ⅲ type V the type I colony highest yield, III-centered, the lowest type II, type V form fruiting bodies are deformed; IV, VI-VII fruiting bodies forming ability. Fruiting bodies of colony forming ability in the fruiting body production in subcultured With the increase of culture generation increased to reduce the higher Following the the cultured middle of several generations fruiting body production. Type I colonies fruiting bodies to maintain the high-yield and high-yield algebraic largest portfolio of mineral elements K2Mg3Ca3Mn3Zn2 Mn and K promote fruiting bodies of high-yield, K, Mg, Ca there is conducive to high to maintain multiple generations. Type II colonies fruiting body production of the highest combination of mineral elements K3Mg2Ca4Mn3Zn2, Zn, Ca, promote the formation of fruiting bodies, Mn depress it. (2) the distribution of various types of colonies, higher production of fruiting bodies of type Ⅰ, Ⅲ the colonies mainly distributed in the first 10 generations, lower production of type II distribution of the first four generations, the formation of fruiting body malformations V-colonies and fruiting body-forming ability of IV, VI-VII colony types are mainly distributed in four generations. 3 different types of colonies algebra different colonies of production, type I colonies algebra longest, followed by the type III colonies, type II algebra shortest. Different mineral elements medium to keep the algebra different effects on different types of colonies, type I colonies algebraic combination of the longest K3Mg1Ca2Mn1Zn3 / 4, K, CA, Zn has played the role of delay degradation, and Mg, Mn will procure its degradation. Type II colonies remain the algebraic maximum the combination is K2Mg4Ca4 Mn2Zn4, type III colonies algebraic combination of the longest K4Mg2Ca2 Mn4Zn3, these five kinds of mineral elements have played the role of delay degradation of type Ⅱ, Ⅲ. Following the generation of the increase in the number of each type of mycelial growth rate showed some leggy trend mycelial growth rate in order to IV GT; VII gt; V gt; VI GT; III GT; II gt; I showed that the mycelial growth slower colonies fruiting body production. Type I colonies mycelial growth slower, less leggy combination K3Mg2Ca2Mn3Zn2, Mn, Zn inhibition of type I colony mycelium leggy; Type II colonies, mycelial growth rate is slower, less leggy mineral elements in combination K2Mg4 Ca3 Mn1 Zn4, inhibition of the largest mineral elements Zn, K, Mn promotes excessive growth of type II colony mycelium. Various types of colonies mycelium dry weight with the subculture of the increase in the number of increases, the dry weight of the order of the various types of colonies for II GT; I GT; IV GT; VII gt; VI GT; III GT; V, show that in colony types form fruiting bodies, the dry weight of mycelium more weight, the lower the yield of fruiting bodies; combination of type I colony dry weight of mycelium lowest K2Mg3Ca2Mn3Zn2, Ca, Mn, type I colony dry weight of mycelium. . For the Ⅱ colonies mycelial dry weight combination of minimum Bad K4Mg3Ca4Mn2Zn4, K, Mg, and Mn influenced. Angle variability results showed that in the presence of corners colony types, various types of colonies of the angular rate of change gradually increases with increasing subculture times. The order of the various types of colonies angle variability as I lt; III LT; II LT; V, show that the higher production of fruiting bodies colonies is more stable, less prone to the corners. The smallest the Ⅰ colonies angle variability of the composition of K3Mg1Ca4Mn3Zn3, wherein K, Ca, Zn, Mn can inhibit the the Ⅰ colony angle variable occurrence; type II colonies angle becomes the minimum rate of the composition K2Mg1Ca4Mn1Zn4 results show that the K, Ca, Zn inhibition of the Ⅱ colony angle becomes occurs. Seven different types of colonies on the different mineral elements medium asexual structure observations suggest that Cordyceps strains asexual multi-type phenomenon, the asexual sporulation characterized divided into two kinds, Paecilomyces and the Cephalosporium type, vegetative characteristics of the original colonies and colonies I also has the structure of two forms of sporulation, sporulation ability, stable colonies traits, higher production of fruiting bodies; Type II, Type III only has Cephalosporium type, sporulation lower colony traits instability, lower fruiting body production; V type only Cephalosporium, its sporulation weak, the abnormal spores to form fruiting bodies deformity; IV, VI-VII sporulation capability, and therefore can not form fruiting bodies. That asexual sporulation colony morphology, fruiting body production has a greater impact, but also has Two sporulation structures are stable colonies, sporulation ability, stable colonies traits, higher production of fruiting bodies, colonies remain algebra longer; only a sporulation structures colony traits unstable and susceptible to degradation. Various types of colony sporulation with the following passage number increased first and decreased following a higher the cultured middle of several generations. Type I colony sporulation medium type III colonies, type II colony lowest. Type I colonies following the culture sporulation combination of mineral elements K3Mg2Ca4Mn3Zn2, including Mn, K sporulation greatest impact; Type II colony subculture the sporulation highest combination K2Mg4Ca4Mn4Zn4, Mg, Zn its impact maximum. Various types of colonies esterase activity in the middle of several generations, enzyme activity were as I GT; III GT; II GT; V gt; IV GT; VII gt; VI. Ⅰ type colonies esterase activity of the highest combination K2Mg3Ca3Mn2Zn2, Ca, K, Mn its greatest impact; the highest combination of type II colony esterase activity K2Mg3Ca4Mn4Zn4, K, Ca, Zn had the greatest impact. 10 colonies of various types of SOD activity is higher in the middle of several generations, SOD activity I GT; III GT; II GT; V gt; IV GT; VII gt; VI, that the degree of aging of the Ⅵ. Ⅰ type colonies SOD activity of the highest combination K3Mg3Ca3Mn3Zn2, Zn, Ca greatest impact; highest activity of type II colonies combination K3Mg3Ca2Mn3Zn2, Zn, Ca, K had the greatest impact. Show that Zn, Ca there is help to improve the Cordyceps SOD activity, thereby increasing the strain of anti-aging ability. 11. Cellulase is a multi-enzyme complex, and the different components of the enzyme activity in the middle of several generations, different colonies carboxymethyl cellulase (CMC), and - glucanase activity order for i GT ; Ⅲ gt; Ⅱ gt; Ⅴ gt; Ⅳ gt; Ⅶ gt; Ⅵ; filter paper activity order as I GT; III GT; II gt; V gt; IV GT; Ⅵ gt; vii showed that the type I colonies cellulase The higher, the different component parts of the enzyme, the production of Cordyceps carboxymethyl cellulase activity is strongest, followed by p-glucanase, the lowest filter paper enzyme. The Ⅰ colonies carboxymethyl cellulase activity combination of the highest Bad K3Mg2Ca2Mn3Zn2, K its largest impact; highest combination of type II colonies carboxymethyl cellulase activity Bad K4Mg3Ca3Mn4Zn2, Mn, K, Ca its greatest impact. Show that K will help improve the ability of of Cordyceps production carboxymethyl cellulase. The Ⅰ colonies beta-glucanase activity of the combination of the highest for K2Mg2Ca2Mn3Zn2, K, Ca its greatest impact; highest combination of type II colonies of p-glucanase enzyme activity K4Mg2Ca4Mn3Zn2 Mg its greatest impact. Type I colonies filter paper activity of the highest combination of carboxymethyl cellulase activity best combination of basically the same, where Mg had the greatest impact; highest combination of type II colony filter paper activity K4Mg2Ca3Mn2Zn4 Mg had the greatest impact. 12 Correlation analysis showed that yield colonies type I, fruiting body production and sporulation, esterase activity, SOD activity, carboxymethyl cellulase activity, p-glucuronidase activity and filter paper activity of correlation were highly significant level of SOD activity in the highest correlation, indicating that the higher type I activity, physiological metabolism, anti-aging ability, stable characteristics, strains, and higher production of fruiting bodies; and mycelial growth speed, dry weight of mycelium angle change rate was negatively correlated, but the correlation was not significant, indicating that the type I colony mycelium is not easy leggy little influence on the fruiting body production. In summary, type I colonies (that is, the original colony type) for excellent colonies, colony characteristics: the colonies color was orange, colony texture is more dense, medium gas endogenous, a concentric pattern, mycelial growth rate is relatively slow, liquid culture mycelium biomass medium, colonies exist corners, but the angle variability than other angle colonies, fruiting bodies of higher yield, the colony algebra longer, the most activity, physiological metabolism, enzyme production ability, The strain of anti-aging ability, Cordyceps excellent colonies; intangible characteristics of two forms of sporulation structure (to be Penicillium and Cephalosporium type). Colony in the middle of several generations fruiting body production, physiological metabolism, so the seed can domesticated generations to ensure high and stable yield. Productive and to keep the algebraic more best combination of mineral elements of K3Mg2Ca4 Mn3Zn2. 5 K in many ways to play a positive role in type I colonies (excellent colony), followed by Ca, again is Mn, the minimal effect of Zn and Mg.