“对于我说，今天是一个很重要的日子。因为我不仅踏进了真正的大学，而且将要研究高科技的生物技术——植物组织培养，也就是现在流行说的植物克隆。

今天，对我来说实在是太重要了。我不仅走进了大学，还学会这么多大学时候才能学到的东西。我想，大学，我已经一步步地向你靠近！”

这是摘自学生在开展个性特长课程学生日记的两句话。有了个性特长课程，也就有了学生的“大学”，学了什么？请看《美国芦荟克隆的研究》

一、选题 Selected[si5lektid] title

克隆人真的会诞生吗？Will the human cloning really be born?

人的智慧能克隆吗？……Can people's intelligence[in5telidV[ns] be cloned?

同学们通过热烈的讨论认为，能自己动手去研究现代的克隆技术最好。于是我们都迫切要求导师能教我们在克隆的技术。王老师按照我们选定的课题，讲述了很多克隆研究的基本知识，尤其是我们目前着手研究的植物克隆技术，王老师讲的更深入一些，又介绍了国内外对植物克隆的研究情况。当介绍到目前华南师大生物系正在克隆美国芦荟的进展情况和发展的前景时。同学们非常兴奋，一致决定要研究这个项目，题目就定为“美国芦荟克隆的研究”。Through[WrU:] warm discussions of the students, we think that it’s best to start and study modern clone technology by ourselves. Then all of us require the tutor[5tju:t[] to teach we the cloning technology[tek5nCl[dVi] urgently[[:dV[nti]. Mr. Wang has told according to the subject that we select a lot about clone ABC, especially[is5peF[li] about the plant clone technology that study, Mr. Wang also introduced the research situation[9sitju:5eiF[n] of the plant cloning both at home and abroad[[5brC:d]. When he introduced the research progress[5pr[Ugres] of plant cloning in Biology Department[di5pB:tm[nt] of South China Normal University, the students are very excited[ik5saitid], and decide to study this project[5prCdVet] together. the topic is determined[di5t[:mind] as“the research of the American aloe cloning”.

我们查找资料知道芦荟是百合科，属多年生植物，原产于南非及马达加斯加岛等地，在世界各地分布很广。芦荟种类有三百多种，而利用价值较高的仅有四、五种，如美国翠叶芦荟、日本木剑芦荟、中华芦荟和开普芦荟等。其中尤以美国翠叶芦荟最具有药用和美容功效，其深加工产品如饮料、保健品、药品和美容化妆品等也早已走进千家万户.We find out that the aloe is lily[5lili] branch, belongs to perennial[p[5renj[l], and originated[[5ridVieit] in South Africa[5Afrik[] and Madagascar[mAd[5gAsk[] island and other places, it can be seen widely in all parts of the world.Aloe has more than 300 kinds, but there are only four or five kinds of high value[5vAlju:]. Such as the American green leaf aloe, Japanese wooden[5wudn] sword[sC:d] aloe, Chinese aloe and opening general aloe, etc.among them the American green leaf aloe has medical[5medik[li] and cosmetic[kCz5metik] efficiency[i5fiF[nsi] most, its deep processed[pr[5ses] product[5prCd[kt] has come into the millions of families[5fAmili] like beverage[5bev[ridV], health products, medicines and cosmetics, etc.

二、研究过程 Research process

根据植物克隆（又叫植物组织培养tissue culture）的技术和有关的要求，我们把研究的过程分为四个阶段：一是MS培养基的制作，二是消毒和灭菌，三是接种，四是培养。According to technology and relevant[5reliv[nt] requests that plant cloning, We divide the course[kC:s] into four stages: 1、MS making of culture, 2、disinfect[9disin5fket] and sterilization[sderilai5zeiFion], 3、inoculate[i5nCkjuleit], 4、train.

（一）培养基制作 Making culture[5kQltF[]

培养基，一个挺陌生的名词。培养基是什么？培养基是为培养植物细胞而设计的，有较高的浓度，保证组织生长所需的矿物质营养和加速愈伤组织生长的物质。总之，培养基里面含有植物生长需要的营养物质。培养基有很多种，我们用哪一种呢？Culture, a very strange[streindV] noun[naun].What is the culture? The culture is designed[di5zaind] for training[5treiniN] the plant cell, there is higher concentration[9kCnsen5treiF[n], organize[5C:g[naiz]and grow necessary[5nesis[ri] mineral[5min[r[l] substance[5sQbst[ns] nutrition[nju:5triF[n] and accelerate[Ak5sel[reit] wounded[wU:nd] to organize material[m[5ti[ri[l] that grow. In a word, culture contains[k[n5tein] nutriment[5nju:trim[nt] that plants need. There are many kinds of culture, which kind do we use?

1、选用什么培养基好？How to select the culture?

课题组在研究培养基时，同学们在网上查找大量的培养基和培养基配方，发现各种培养基都有各自的特点：MS培养基适合固体培养，无机盐的数量和比例适中，能满足大多数植物的生长。B6培养基含铵较多，对某些植物生长有好处。N6培养基主要用于花的培养。White 培养基是含浓度低的无机盐，主要用于胚的培养。我们采访了华南师大的植物克隆专家，他们也认为MS培养基比较适合美国芦荟的生长，因此，课题组成员决定采用MS培养基。The seminar[5seminB:],at the time of studying the culture, the classmates find out on the net that a large amount[[5maut] of culture and culture prescription[pri5skripF[n],and find various[5vZ[ri[s] kinds of culture have their own characteristics[9kArikt[5ristik]:

MS culture suitable for solid train,quantity[5kwCntiti] and proportion[pr[5pC:F[n] of inorganic[9inC:5gAnik无机的] salts moderate[5mCd[rit],can satisfy['sAtisfai] most growth of plant.

2、培养基的具体制作：The processpr[5ses] of making the culture

（1）从母液中取出所需要的大量元素、微量元素、铁盐、维生素以及植物激素等，放于烧杯中；Take out a large number of elements[e5l[m[nts], trace[treis] element['elim[nt] ,molysite['m[Ulisait],vitamin['vait[min] and plant hormone['hC:m[Un]needed,etc.from the mother liquor['lig[] and put in the beaker['bI:k[]

（2）煮琼脂，加热并要不断搅拌，琼脂煮沸约十几分钟后便会溶解，呈透明状；Boil agar['eigB:], heat it while stir[st[:]it constantly[5kCnst[ntli], after boiling the agar for over ten minutes, it will dissolve[di'zClv] and be transparent[trAs'pZ[r[nt透明]

（3）琼脂溶解后，加入从（1）中所取出的各种药物及糖，同时要不断搅动培养基使其混合均匀；After dissolving[di'zClviN] agar, put in various['vZ[ri[s] kinds of medicine and candy taken out from (1),you should stir[st[:]constantly to make the culture mixed.

（4）用“pH计”或“pH试纸”测PH值。用浓度为1摩尔氢氧化钠或1摩尔盐酸将培养基调至适合的PH值，通常为PH=5.8；With “pH meter”or“pH test paper”examine hydrogen['haidr[UdV[n]ion['ai[n].Using the concentration[9kCnsen5treiF[n] of 1 Mor sodium[5s[Udj[m]hydroxide['haidrksaid]or 1 Mor hydrochloric[9haidr[U5klC:rik] acid['Asid] to mix the culture train until the pH suits, the pH is usually 5.8

（5）将调配好的培养基用漏斗或分装器分装到培养用的玻璃瓶内。这些玻璃器皿要事先清洗干净、烤干。琼脂约在40℃时才凝固。 Allocate['Al[Ukeit] culture and put it into vial['vai[l] with funnel['fQn[l]or partial['pB:F[l] device. These glass ware[wZ[陶器] should be washed clean,dry in advance[[d5va:ns]. Agar just solidified[s['lidifai] on the about 40℃

（6）将分装好的培养基容器盖上盖，标上记号准备高压灭菌。这样，就把培养基做好了。cover the each container[k[n5tein[] with the culture and mark[ma:k]them for high-handed sterilization['sterilai'zeiFion]. In this way, we can finish making the culture.

（二）灭菌 Sterilize['sterilaiz]大家都知道，培养基是最有利于植物、细菌和真菌繁殖的物质，如果在实验的过程中落入细菌或真菌孢子，整个实验就会失败。如何防污染？我们研究和实践后认为要注意以下3点：Everybody knows,the culture is the material[m[5ti[ri[l] helping the plant,bacterium[bAk5ti[ri[m细菌] and fungi['fQNdVai] to breed[bri:d]most.If course in the experiment decline[di'klain] into bacteria[bAk5ti[ri[m] or the fungi['fQNdVai]spore[spC:], the whole[h[Ul] will fail[feil] .how to prevent[pri'vent] the experiment from pollution[p[5lU:F[n污染]? We think that we should pay attention to the three aspects['Aspekt方面] after we tried the experiment:

1、保持实验室的绝对干净。实验前要对实验室进行紫外线灯灭菌20分钟，如是超净工作台则开机过滤空气15-20分钟。Keep the lab[lAb]clean. Before experiment we should sterilize['sterilaiz]with ultraviolet['Qltr[5vai[lit]lamp for 20 minutes,if there is an ultra['Qltr[] workingbench,we should start filter ['filt[过滤]15-20 minutes.

2、器具消毒灭菌。首先用清水洗净，然后用中性洗涤剂洗，再用清水冲洗数次，接着用蒸馏水洗2-3次，最后就是晾干备用，烘干灭菌。将洗净的器具置于烘干箱，150℃烘2小时，即可达到灭菌目的。The utensil[ju:5tensl器具]disinfects[9disn'fekt] and sterilizes['sterilaiz].Clean with distilled[dis5tild]water at first,wash with neutral['nju:tr[l 中性]detergent[di't[dV[nt清洁剂],then wash with the distilled[dis5tild] water for several times again.at last, dry sterilization[sterilai'zeiF[n] for future use. Put clean utensil[ju:5tensl]on case of drying,bake[beik] 2 hours at 150℃,that's done.

3、材料消毒。美国芦荟外植体用漂白粉消毒，浓度一般为5-10%，也可以用饱和溶液。消毒时间为10-20分钟。The material[m[5ti[ri[l] disinfects[9disn'fekt].disinfect the American aloe plant body with the bleaching['bli:tFiN]powder,the concentration[9kCnsen5treiF[n] is generally['dVen[r[li]5-10%, also we can use the saturated['sAtF[reitid]solution[s[5lju:F[n]. The time of disinfecting[9disn'fektiN] is as 10-20 minutes.

4、培养基的灭菌。MS培养基在120摄氏度，压力保持在1.1千克/平方厘米，20分钟后即可高压灭菌。Sterilization['sterilai'zeiFion]of the culture.MS culture at120℃at 1.1 kilograms['kil[grAm]/square[skwZ[]centimetre['senti9mi:t[r]pressure['preF[(r)],20 minute heat sterilization['sterilai'zeiFion].

5、规范的无菌操作法。组培技术是一种无菌技术，除了保持工作室清洁无菌外，工作人员也要注意不带细菌，要剪除指甲，穿工作服，带工作帽和口罩。操作前先用肥皂洗手，操作时用70%酒精擦手灭菌。接种时，需在近火焰上方打开培养基瓶盖子，使瓶倾斜，以免空气中微生物落如瓶内。接种完毕，应立即盖好瓶口，用包头纸包扎紧。Normal aseptic[A5septik]manipulation[m[9nipju5leiF[n].Culture-developing is asepsis[A5sepsis 无菌]technology, besides keeping the workroom clean and asepsis[A5sepsis], members should pay attention to not taking either, should wipe[waip] out the nails[neil],wear the work clothes,take the job caps and gauze[gC:z] masks[ma:sk].Wash hands with soap[s[Up]first before operating,wipe[waip] hand sterilization['sterilai'zeiFion] with 70% of the alcohol['Alk[hCl酒精]when operating.When inoculating[i5nCkjuleitiN],open culture bottle shell[sel] above nearly flame[fleim],make the bottle slope[sl[Up倾斜],In case[keis]that the microorganism[maikr[U5C:g[nizm] in the air falls[fC:l] in the bottle. After Inoculate[i5nCkjuleit],we should cover bottleneck properly['prCp[li]at once, wrap[rAp]up with clean paper.

（三）接种inoculation[i5nCkjuleiFion]接种是植物克隆的重要的操作阶段，如何做好这个阶段的工作？我们研究和实践后认为，接种过程可以分为以下6个步骤。It is the important operation stage[steidV] that a plant clones to inoculate[i5nCkjuleit], how to do? After we research and practice[5prAktis],we think that inoculating[i5nCkjuleitiN]can be divided into the following 6 means['mi:s].

1、把接种所需要的物品和培养基在超净工作台上摆好。Put the article and culture needed on the ultra['Qltr[] inoculating.

2、接种室进行紫外线灯灭菌20分钟，超净工作台同时开机20分钟。sterilize['sterilaiz]the inoculate[i5nCkjuleit]room with ultraviolet['Qltr[5vai[lit]lamp for 20 minutes, start the workingbench 20 minutes at the same time.

3、进入接种室前更换衣物并把手用70%的酒精擦洗。before entering, change the clothes and scrub[skrQb]hands with 70% of the alcohol in front of the room.

4、接种工具在酒精灯上烧灼灭菌。Inoculate[i5nCkjuleit]tools and burn sterilization on the alcohol['Alk[hCl]burner .

5、切割美国芦荟外植体并在酒精灯上方接入培养基中。Cut American aloe plant body and insert culture above alcohol['Alk[hCl]burner.

6、将瓶口和瓶盖在酒精灯上转动消毒，盖上瓶盖。Turn and disinfect[9disn'fekt] bottleneck and bottle lid(盖子) at alcohol['Alk[hCl] burner, cover the bottle lid.

（四）培养 Training接种之后，进行固体培养。培养的条件:是After inoculating[i5nCkjuleitiN],carry on the solid to train.The condition trained is:

温度：25℃。Temperature: 25 degrees Centigrade.

（五）结论 conclusio[k[n`klu:V[n]通过本次研究实验，我们做了8瓶美国芦荟外植体的培养，到目前为止，没有出现一瓶受到微生物感染，而且生长正常。因此我们可以得出以下结论：Through this research experiment,we make 8 bottles of the American aloe plant cultivation[9kQlti`veiF[n],Up till now,not presenting[pri`zent]one bottle infected[in'fektid]by microorganism[maikr[U5C:g[nizm],and it grows normally[`nC:m[li].So we can draw[drC:] the following conclusions[k[n`klu:V[n]:

我们的实验操作已经达到无菌的要求。Our experiment meet the requirement asepsis[A5sepsis 无菌].

Because time is short,we have the following work to do and study,for instance['inst[ns]: How to improve[im'prU:v] Ms culture,make it to be easy to adapt[[5dApt]to the American aloe.The American aloe has not been trained completely[k[m`pli:tli].

1、为什么克隆的对象要选择美国芦荟呢？Why will we choose[tFU:z]the American aloe cloning?　

因为芦荟这种植物在现代社会越来越流行，我们查找资料知道，芦荟种类有三百多种，而利用价值较高的仅有四、五种，其中又以美国翠叶芦荟最具有药用和美容功效，其深加工产品如饮料、保健品、药品和美容化妆品等也早已走进千家万户。Because aloe are becoming more and more popular[5pCpju:l[]in the modern society[s[5sai[ti],Aloe has more than 300 kinds, but there are only four or five kinds of high value[5vAlju:]. Such as the American green leaf aloe, Japanese wooden[5wudn] sword[sC:d] aloe, Chinese aloe and opening general aloe, etc.among them the American green leaf aloe has medical[5medik[li] and cosmetic[kCz5metik] efficiency[i5fiF[nsi] most, its deep processed[pr[5ses] product[5prCd[kt] has come into the millions of families[5fAmili] like beverage[5bev[ridV], health products, medicines and cosmetics, etc.

一共是四个阶段：一是MS培养基的制作，二是消毒和灭菌，三是接种，四是培养。It has four stages:1、MS making of culture, 2、disinfect[9disin5fket] and sterilization[sderilai5zeiFion], 3、inoculate[i5nCkjuleit], 4、train.

MS培养基适合固体培养，无机盐的数量和比例适中，能满足大多数植物的生长。我们采访了华南师大的植物克隆专家，他们也认为MS培养基比较适合美国芦荟的生长，因此，课题组成员决定采用MS培养基。MS culture suitable for solid train,quantity[5kwCntiti] and proportion[pr[5pC:F[n] of inorganic[9inC:5gAnik无机的] salts moderate[5mCd[rit],can satisfy['sAtisfai] most growth of plant.

培养基是为培养植物细胞而设计的，有较高的浓度，保证组织生长所需的矿物质营养和加速愈伤组织生长的物质。The culture is designed[di5zaind] for training[5treiniN] the plant cell, there is higher concentration[9kCnsen5treiF[n], organize[5C:g[naiz]and grow necessary[5nesis[ri] mineral[5min[r[l] substance[5sQbst[ns] nutrition[nju:5triF[n] and accelerate[Ak5sel[reit] wounded[wU:nd] to organize material[m[5ti[ri[l] that grow.

灭菌。因为培养基是最有利于植物、细菌和真菌繁殖的物质，如果在实验的过程中落入细菌或真菌孢子，整个实验就会失败。Sterilization[sderilai5zeiFion].Because the culture is the material[m[5ti[ri[l] helping the plant,bacterium[bAk5ti[ri[m]and fungi['fQNdVai]to breed[bri;d]most,If in the experiment decline into bacteria[bAk5ti[ri[]or the fungi['fQNdVai]spore[spC:],the whole experiment will fail.

组培技术是一种无菌技术，除了保持工作室清洁无菌外，工作人员也要注意不带细菌，要剪除指甲，穿工作服，带工作帽和口罩。操作前先用肥皂洗手，操作时用70%酒精擦手灭菌。接种时，需在近火焰上方打开培养基瓶盖子，使瓶倾斜，以免空气中微生物落如瓶内。接种完毕，应立即盖好瓶口，用包头纸包扎紧。Culture-developing is asepsis[A5sepsis 无菌]technology, besides keeping the workroom clean and asepsis[A5sepsis],members should wipe[waip] out the nails[neil],wear the work clothes,take the job caps and gauze[gC:z] masks[ma:sk].Wash hands with soap[s[Up]first before operating,wipe[waip] hand sterilization['sterilai'zeiFion] with 70% of the alcohol['Alk[hCl酒精]when operating.When inoculating[i5nCkjuleitiN],open culture bottle shell[sel] above nearly flame[fleim],make the bottle slope[sl[Up倾斜],In case[keis]that the microorganism[maikr[U5C:g[nizm] in the air falls[fC:l] in the bottle. After Inoculate[i5nCkjuleit],we should cover bottleneck properly['prCp[li]at once, wrap[rAp]up with clean paper.

　7、如何进行器具消毒灭菌？How to carry on utensil disinfect and sterilization ?

首先用清水洗净，然后用中性洗涤剂洗，再用清水冲洗数次，接着用蒸馏水洗2-3次，最后就是晾干备用，烘干灭菌。将洗净的器具置于烘干箱，150℃烘2小时，即可达到灭菌目的。The utensil[ju:5tensl器具]disinfects[9disn'fekt] and sterilizes['sterilaiz].Clean with distilled[dis5tild]water at first,wash with neutral['nju:tr[l 中性]detergent[di't[dV[nt清洁剂],then wash with the distilled[dis5tild] water for several times again.at last, dry sterilization[sterilai'zeiF[n] for future use. Put clean utensil[ju:5tensl]on case of drying,bake[beik] 2 hours at 150℃,that's done.

美国芦荟外植体用漂白粉消毒，浓度一般为5-10%，也可以用饱和溶液。消毒时间为10-20分钟。disinfect the American aloe plant body with the bleaching['bli:tFiN]powder,the concentration[9kCnsen5treiF[n] is generally['dVen[r[li]5-10%, also we can use the saturated['sAtF[reitid]solution[s[5lju:F[n]. The time of disinfecting[9disn'fektiN] is as 10-20 minutes.

9、美国芦荟要在什么温度下培养？Under what temperature does American aloe need to train? 25℃.

10、在整个研究过程中，你们遇到了哪些困难？During the whole research,what difficulties[`difik[lti]have you met?

一开始研究的时候，我们对生物还处在初级阶段，物理和化学都没有接触，所以概念理解起来比较困难，操作也十分的生疏。进度很慢。At the beginning of research, we know a little about living things, and know little about physics and chemistry,So it is more difficult to understand in concept[5kCnsept概念],it is new to operate. The progress is very slow.

我们在课外花多一点时间对知识进行理解，不懂就问，每一个操作反复做几遍，慢慢就熟练起来了。We spent more time getting to know the knowledge, and question whenever[wen`ev[]needs.Repeat each operation,then you’ll be skillful[5skilful].

我们学到了进行科学研究的严谨性，还学会了如何对问题进行分析。We learnt that scientific[sai[n`tifik]research needs preciseness[pri`saisnis] and how to analyze[5An[laiz].

13、进行这样的研究，对你们的学习有影响吗？What’s the impact[`impAkt]on your study while doing such research?

没有影响，反而增添了我们对学习的兴趣，培养了我们在学习上坚持不懈的精神。这对于我们以后的学业是很有帮助的。No negative[5neg[tiv]impact[5impAkt],instead,it interests and inspires[in5spai[]us.it is very helpful for our the future study.

14、除此之外，你们还有什么特别深的感受？In addition[[`diF[n],do you have any very deep impression[im`preF[n]?

进行这样的研究，任务很艰巨，但很快乐。其实做事情要注重的是过程，但是当我们的成果诞生了之后，心里的欢乐是无与伦比的，也特别的有成就感。Carrying on such research, the task is very arduous[5B:dju[s], but very happy. We focus[5f[Uk[s] on the procedures[pr[5si:dV[] and when we have the achievement[[5tFi:vm[nt], we are so satisfied and proud[praud].

15、你们对你们的研究有什么展望吗？What do you expect[iks5pekt] for your research?

我感觉到我们的研究应该开始一个更新的阶段。在这个阶段里，我们要做更深入的探索，相信会有更大的成果。I feel that our research should begin a new stage.During this stage, we want to make deeper exploration[9eksplC:5reiF[n],believe that there are heavier achievements.