Study On The Technique Of Clone Culture In Gametophyte Laminaria

This article mainly discusses the large-scaled cultural conditions of clone in Gametophyte Laminaria .It studies the proper temperature, light time, light intensity, the consumption of nitrogen and phosphorus, the cultural state and inoculating density on the growth of gametophytic clone, the experiment of large-scaled culture had been done under comparatively proper conditions, in the same time, we studied the growth rule of gametophytic clone in the cultural process. We want to supply theoretic base and technical sustentation for large-scaled culture of clone, offer premise condition for the translation from traditional rearing manner to clone breeding method. The results indicate that:Temperature is one of the most important environmental factors impacting the growth of clone . It has the fastest growth speed under10-15℃. In our experiment, the clone we used almost all died under 25℃. In the same condition, the female and male clone cell that have mostly same number have close growth speed.Light plays very important role in the growth of Gametophyte clone. Light time and intensity affect its growth directly. When the light time is 24h and intensity is 1500-2500Lux, the growth speed of clone is fast.The results of the consumption of nutrition indicate that when the density of clone is lower ,changing the cultural fluid once a week, N6ppm and Plppm is enough; when the density of clone is higher, N-10ppm and P-2ppm is sufficient.In the condition of suspending culture, the gametophytic clone has much higher instantaneous growth rate than stilling culture. Suspending culture is fit for productive cultivation.The growth speed of clone is different in various inoculating density. The experiment result indicates: Lower is inoculating density, faster the speed of growth is. If conditions permit, using more utensils will improve the general yield of clone.In the course of culture, the growth disciplinarians of clone are: in the initial stage, when the density is lower, its instantaneous growth rate increases gradually; With the continuity of culture and increasing of density, the growth speed declines.In jars of 20L, the inoculating density is 0.5g/L, providing proper conditions, experiments of five repeating groups were conducted. According to the average values, the curve of fresh weightiness can be drawn. The formula, y=-0.1118+1.3108x+0.085x2-0.0006x3 (R2=0.9985), was gained by regression analysis and it can be used to estimate products of clone (y is fresh weightiness, x is time) .The results of large-scaled culture show that 48g clone crushed by tissue disintegrator in the course of initial period and its inoculating density is 0.35g/L, a month later ,the clone departed directly and its density is 1.5g/L, the highest density of each jar could be obtained 410g, the growth speed mostly doubled per week. Calculating according to this , a breeding plant of 4 hundred millions throughput (medium scale) needs 36kg clone, which can be produced in two months and only requires about 100 jars of 20L. It is obvious that the clone which breeding production needs can be gained easily under manpower in chamber utensils.