The Preparation Of Reproductive Cells Of Ulva Pertusa And The Observation Of Its Early Growth

Ulva pertusa is a common green alga of the coastal our country, it widelydistribute in coastal of the West Pacific Ocean. It has broad application prospect infood, pharmaceutical, environmental protection. But at present, the yield and qualityof Ulva pertusa are difficult to meet the business requirement at home. The selectionand multiplication of its varieties are the key to solve the problem. But there has notformed mature artificial breeding technology, not to mention large scale farming. Atpresent, domestic mainly adopt collecting spores and enzymatic hydrolysis to prepareartificial breeding source of algae. This article studies the influencing mechanism onmaturity of Ulva pertusa by illumination intensity、temperature and salinity andoptimized the enzymatic hydrolysis conditions of protoplast and then observes theearly growth of Ulva pertusa, providing theoretical basis for breeding and technicalsupport. The experimental results are as follows：1The influencing mechanism on maturity of Ulva pertusa by illuminationintensity、temperature and salinity1.1The effect of illumination intensity and temperature on maturity of Ulva pertusaUlva pertusa was collectted in the open. To probe into the appropriateconditions of maturity and discharge through the cross experiment of temperature andlight. The result showed that,30days after inducing, Ulva pertusa discharged onconditions of20℃,16000Lux.in addition, the conditions of25℃,4000Lux nd25℃,8000Lux also made for Ulva pertusa to mature.1.2The effect of salinity on maturity of Ulva pertusaReduce the salinity of the seawater to get different salinity culture solution of0,5,10,15,20,25and the control is natural salinity. Observe the effect on maturity ofUlva frond which comes from the different salinity. And it can provide the theoreticalbasis for desalination domesticated of Ulva pertusa. The result is, Ulva pertusa whichthe salinity is10, there are clearly particulate matters showing that Ulva frond start tomature. Ulva pertusa which the salinity is15,20and25, there are more particulatematters, showing that the maturity is higher than former. Among them, the salinity is10, there are the most particulate matters which has the highest maturity.2.Optimization of enzymatic hydrolysis conditions of protoplast of Ulva pertusaTo determine the optimal enzyme digestion conditions, this articalrespectively does research about the effect on the protoplast harvest of Ulva pertusa,which comes from the combination, mixture ratio of the enzyme, the type andconcentration of the enzyme-benign solvent, enzyme digestion time, enzyme solutiontemperature and the pH of the enzyme solution environment. The best enzymolysisconditions are：the optimal enzyme mixture is0.5%macerozyme,2%pectinase,2%cellulase, the optimal enzyme solvent is mannitol of0.7mol/L, the optimal pH is5.7,the optimal enzymolysis time is3hours and the optimal enzymolysis temperature is28℃.3Observation of early growth of Ulva pertusa3.1Observation of early growth of gamete from wild Ulva pertusaMature Ulva pertusa were collected in the outdoors. It discharged gametesnaturally, which were identified through KI-I2staining. The gamete developed asfollows: first cell division mainly happened in the vertical direction of the gamete, andthen the gamete developed into a cell stick after a week. Cell division in the horizontaldirection occurred on the ninth day. The rhizoid formatted on the tenth day. After acouple of weeks, filament seeding formatted. Then the next was thallus.3.2Observation of the early growth of single cell released by rotten wild Ulva pertusaSingle cells could be released when Ulva pertusa was rotten. These cellswere larger than spores and gametes which were discharged by Ulva pertusa. The cells could grow into protonema by means of cell division.This may be thecharacteristic reproductive strategy.3.3Observation of early growth of clonal propagation of Ulva pertusa from JapanThe explant which came from clonal propagation of Ulva pertusa fromJapan could crack into different shape of multicellulars. These multicellulars had twodevelopment patterns. One of the pattern is developing into sporangium anddischarged spores after mature. The other is discharging cells directly. The spores andcells could divide and grow into protonema.