克隆模式哪个游戏最先出的

目前来看，好像是英雄联盟

英雄联盟克隆模式上线时间

One for All is a limited time game mode in League of Legends Players battle in a 5v5 match in Summoner's Rift map similar to Classic mode, with the only difference being that all champions on each team are the same The mode was first made available from 2013-11-02 to 2013-12-02 It is occasionally relaunched to align with April Fools' Day

源自>

One of all模式（可译成克隆模式）是一个有限的时间游戏模式在英雄联盟。玩家在召唤师裂谷地图中的5v5比赛中战斗，类似于经典模式，唯一的区别是每个团队的所有冠军都是相同的。该模式首次于2013年11月2日至2013年12月2日提供。它偶尔会重新推出，以配合愚人节。（译文源自百度翻译，仅供参考。）

300英雄

网上说是2013年12月11日开始。

什么是克隆
克隆是英语单词clone的音译，clone源于希腊文klon，原意是指幼苗或嫩枝，以无性繁殖或营养繁殖的方式培育植物，如杆插和嫁接。
如今，克隆是指生物体通过体细胞进行的无性繁殖，以及由无性繁殖形成的基因型完全相同的后代个体组成的种群。克隆也可以理解为复制、拷贝，就是从原型中产生出同样的复制品，它的外表及遗传基因与原型完全相同。
1997年 2月，绵羊“多利”诞生的消息披露，立即引起全世界的关注，这头由英国生物学家通过克隆技术培育的克隆绵羊，意味着人类可以利用动物身上的一个体细胞，产生出与这个动物完全相同的生命体，打破了千古不变的自然规律。
如何评价克隆技术？
无论“雷里安”如何狡辩、美化自己的行为，世界许多著名科学家的看法十分相近：“雷里安”进行克隆人实验没有任何科学目的，一句话，并非为了科学进步。
不少科学家认为，在评论克隆人这个事件时，重要的是应该先弄清楚：人类到底需不需要克隆人？
莫斯科谢琴诺夫医学院遗传学教研室阿利阿萨诺夫教授评论道，技术和工艺方面的可能性大大超过了我们对“人类需要什么”的理解。
克隆人赞同者的论据是，该技术能够帮助不孕者拥有自己的后代。
实际上，这个要求可以通过其它更安全更有效的途径来满足。因此可以断定，利用克隆技术进行传宗接代只是借口，克隆人实验背后隐藏着非科学的商业目的。
阿萨诺夫教授认为，眼下，克隆人没有任何前景，也没有任何意义。值得指出的是，现在没有人能够预言克隆人会产生什么后果，因此现在进行克隆人实验是不道德的。
修理病变器官是克隆的未来
阿萨诺夫教授说，俄罗斯科学界坚信，克隆技术的未来应该是在内科疗法中的应用，即“内科疗法克隆”。不过，现存的问题是，该术语在表达上还极其不准确。
从本质上讲，“内科疗法克隆”是建立移植细胞材料的方法，在意义上与现在所指的克隆没有共同之处，它是一种能够培养健康器官的细胞工艺技术，利用该技术可以部分或全部替换病变器官。
根据阿萨诺夫教授的解释，现在科学家刚刚触及到人体体内所发生的内部过程这个问题，只略知皮毛。科学家前不久解读了人类基因图谱，但还不能很好地应用所得到的知识来揭开人体奥秘。为此，科学家还要进行若干年的深入研究，才能完善并掌握克隆技术。
现在的克隆，百分之九十九将是丑八怪
阿萨诺夫教授说，俄罗斯科学家已经不止一次发出警告，克隆试验所得的产物99％是丑八怪。
他们的例证为：著名的克隆羊多利是经过300次失败后才获得的。遗憾的是，多利并不是一只健康的小羊，它患有关节炎等疾病，而且出现早衰病征。另外，在其它所有克隆动物身上都发现了各种发育畸形。包括阿萨诺夫教授在内的俄罗斯科学家认为，在这种情况下进行克隆人实验，至少是一种极不负责任的做法。克隆人的一生将是一场噩梦，到30岁时，他们将成为苍老之人。
什么东西可以科隆
应该说有生命的都可以克隆
现在已经克隆什么
蛙: 1962年，未成功
鲤鱼: 1963年，中国科学家童第周早在1963年就通过将一只雄性鲤鱼DNA插入来自雌性鲤鱼的卵成功克隆了一只雌性鲤鱼，比多利羊的克隆早了33年。但由于相关论文是发表在一本中文科学期刊，并没有翻译成英文，所以并不为国际上所知晓。（源自：PBS)
绵羊: 1996年，多利（Dolly）
猕猴: 2000年1月，Tetra，雌性
猪: 2000年3月，5只苏格兰PPL小猪；8月，Xena，雌性
牛: 2001年，Alpha和Beta，雄性
猫: 2001年底，CopyCat（CC），雌性
小鼠: 2002年
兔: 2003年3-4月分别在法国和朝鲜独立地实现；
骡: 2003年5月，爱达荷Gem，雄性；6月，犹他先锋，雄性
鹿: 2003年，Dewey
马: 2003年，Prometea，雌性
狗: 2005年，韩国首尔大学实验队，史纳比
尽管克隆研究取得了很大进展，目前克隆的成功率还是相当低的：多利出生之前研究人员经历了276次失败的尝试；70只小牛的出生则是在9000次尝试后才获得成功，并且其中的三分之一在幼年时就死了；Prometea 也是花费了328次尝试才成功出生。 而对于某些物种，例如猫和猩猩，目前还没有成功克隆的报道。而狗的克隆实验，也是经过数百次反覆试验再得来的成果。
多利出生后的年龄检测表明其出生的时候就上了年纪。她6岁的时候就得了一般老年时才得的关节炎。这样的衰老被认为是端粒的磨损造成的。端粒是染色体位于末端的。随着细胞分裂，端粒在复制过程中不断磨损，这通常认为是衰老的一个原因。然而，研究人员在克隆成功牛后却发现它们实际上更年轻。分析它们的端粒表明它们不仅是回到了出生的长度，而且比一般出生时候的端粒更长。这意味着它们可以比一般的牛有更长的寿命，但是由于过度生长，它们中的很多都过早夭折了。研究人员相信相关的研究最终可以用来改变人类的寿命。
克隆人
由于伦理和现实上可能的后果，克隆人一直是一个充满争议的话题。许多人认为克隆人的尝试是不道德的，但有些科学家公开宣称尝试克隆人。一些团体声称他们正在进行克隆人研究或者已经克隆出了人，但是没有独立的消息来源证实。
回答者：小璋 - 经理 四级 3-22 19:32
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克隆，是英语“clone”一词的译音。作名词使用时，表示从一个共同祖先无性繁殖下来的一群遗传上一致的DNA分子、细胞或个体所组成的生命群体。作动词使用时，是指这种无性繁殖的过程。在重组DNA技术中，基因克隆是将特定基因或基因组，插入到能够自主复制的DNA载体上，而引入到寄主细胞中进行增殖的 *** 作，从而为遗传上同一的生物品系的大量繁殖和生长提供了有效途径。克隆技术的问世，必将对人类社会的发展产生深远的影响。

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回答者：小洁婷 - 试用期 一级 3-22 19:33
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摘要：
本文综合性阐述了我国当前果苗生产存在的问题与限制因素，分析了国外种苗生产的趋势，提出了利用新型的育苗技术----植物非试管快繁技术，来实现种苗生产工厂化、自动化、产业化的生产模式，结合生产科研实践概括性地论述了快繁技术在果树种苗培育上的各方面应用与优势，并指出了各种不同果树种类的快繁技术要点，展示了该技术在果树种苗快繁上运用的广阔前景与市场空间。
关键词：植物非试管快繁技术，果树，苗木，自根苗，脱毒苗，工厂化，草地果园
引言
果树生产是我国农业生产中一大重要的产业，是许多地区农民致富农村经济发展的主要经济来源，是项传统的支柱产业。它有悠久的栽培历史，有丰富的管理经验。在种苗的繁殖，到生产栽培管理还有收获加工，形成了产前、产中、产后的产业链。但分析我国目前果业的发展情况及与国际发达国家相比，我们在产前与产后两大环节相对薄弱，这也是影响当前我国果业在国际上市场上竞争力不强的主要因子。如产前的品种选育到快速扩繁直至生产上品种结构的调整，都显得非常薄弱，在新品种的推广进程上显得极为缓慢，在繁殖的手段上显得极为传统与落后，与发达国家的工厂化快速育苗相比距离相差甚大〔1〕。我国目前的状况是种苗培育的业主零散，甚至有些地方还是自育自栽，还有技术上落后，培育的种苗符合壮苗标准较少，没有形成规模化产业化的育苗趋势，这主要也育苗技术的手段方法落后外，还与靠天育苗的生产力状况有关，如发达国家的设施大棚育苗、穴盘容器育苗及无毒化的脱毒育苗在我国应还很少。这些因子都成为当前果树苗木产业化工厂化形成难的主要限制因子，针对这些问题，我国科研生产单位也相继投入研究与引进先进的育苗技术，引进国外的生产技术流程与相应的育苗设施，如组培脱毒，设施栽培，容器基质育苗等技术，但效果不是很理想，一是投入大一般的育苗专业户难以承受，二是技术的 *** 作还需有专业基础的支持。所以尽管不断地引进发达国家技术，但真正消化吸收用于生产的甚少。针对这些问题，浙江省丽水市农科所农业智能化快繁中心涉足了这个领域的研究，在引进国外先进发达技术同时结合我国国情与生产力状况，开发了一项新型的育苗技术叫植物非试管快繁技术，现就针对这项技术在果树育苗上的运用作些介绍，让更多的生产科研者能掌握了解与运用这项技术，利用它来培育出大量能用于当前产业化发展的价低质优的商品苗。
植物非试管快繁技术是一项全新的育苗技术
植物非试管快繁技术是基于传统扦插育苗与组培技术基础上面发展起来的一项新技术，它是利用计算机环境控制手段为植物离体材料创造最佳的温光气热环境，让植物的根原基快速表达，让根系充分发育的技术，同时它又结合了营养液无土壮苗技术，实现离体材料的快速增殖与多代循环相结合达到种苗数量几何级倍增的技术。运用它可以使果树的一叶一芽离体材料在年周期内实现百倍甚至千万倍增殖扩繁的技术，这对于促进一个新品种的快速推广，推动品种产业结构调整来说意义重大。现把它的一些特点作些简要介绍。（1）利用计算机环控技术后可以实现果树种苗的周年快繁，不像传统育苗那样受季节的限制。（2）利用快繁技术可以为生产提供大量遗传基因稳定，性状一致的无性苗。（3）可以使一些在常规扦插技术下根本不能生根的品种，快速生根成苗。（4）可以节省大量传统育苗下育砧与嫁接的繁琐 *** 作达到降低成本的目的。（5）可以实现母本材料在年周期内几何级倍增，大大加快新品种的推广速度。（6）快 快苗对于果树的矮化密植及早产丰产具有很大生理促进作用。（7）可以实现周年移栽周年建园的技术效果。（8）如果结合隔离快繁还可大大降低脱毒苗的生产培育成本，大大推进脱毒苗在生产上的运用与普及。
如一些平常技术下难生根的桃梅李杏樱桃板栗杨梅枇杷等，可以利用快繁技术让其快速生根，培育时，只需截取优良品种的一个枝段或者一叶一芽，就能实现短期内快速生根成苗。这些品种是在常规下难以实现的，那么究竟是什么因素促进一些传统与常规下不能扦插成活或组培不能成功的果树品种，在快繁技术下就能让其生根成苗呢？关键在于育苗气候环境的优化，以及结合了促进快速生根的各项综合技术措施。如一改原来的大田扦插或者室内组培变为以无机基质为载体，以营养液激素为促进为补充的技术改进；一改传统组培的密闭环境为大田苗床的开放环境，一改传统的自养半自养生根过程为全光照全自养的生根过程。这些技术措施的改进为果树离体材料的生根构建了最佳的生理模式。如扦插育苗，在无叶的硬枝条件下，插于土壤中让其生根，就会遇到环境温度变化的不协调性，大多先萌芽后生根，使枝内的营养大多耗于芽的萌发与生长，使根系的营养得不到最大化供给；另外，在土壤环境，常遇切口的病菌感染与水分过湿过干造成的生根阻碍。在有叶片时的带叶扦插，常为了避免叶片过份水分蒸腾而导致水份供求失衡干枯，而采用剪去叶片方法，而造成光合营养不足，因带叶的果树枝段，生根的营养与激素需求主要靠叶片光合作用提供。但如果采用蔗阴以减免蒸发，又会造成光照不足，光合作用不充分，从而影响生根，所以针对果树品种来说大多属于难生根的品种，自然在传统扦插育苗下就难以实现。另外，组培技术也是一样，通过多年研究还是不能解决许多果树的生根培养与炼苗这个问题，许多品种既使能完成芽的增殖培养，但生根培养又遇到难生根的障碍，甚至生根后，炼苗移栽环节还要失败。关键问题，组培过程是个异养与密闭环境下培养的过程，所培育出来的苗木光合作用、呼吸作用常存不正常现象[2]，同时密闭试管空间因处于高湿低氧低二氧化碳环境培育而成的苗木，对外界适应性极差，所以炼苗驯化移栽成为组培是否成功的一个主要生产问题，另外组培因化学药剂的诱导常使种苗发生遗传变异而影响苗木的纯正性。而非试管快繁是在全光开放无机基质的环境下利用材料叶片自身的光合作用能力而启动生根基因，达到成苗目的的技术过程。非试管快繁技术通常利用珍珠岩蛭石等疏松透气不含糖的无机基质为载体，可避免土壤有机物与组培加糖而引致的病菌滋生感染问题，在这种环境下既使细菌真菌进入苗床也不会滋生蔓延。同时计算机环控技术的结合为离体材料生根，光合作用创造模拟出最佳的环境，达到光合自养过程的最大化[3]，切口生根部位环境的最优化。还结合了二氧化碳强制供应技术，使单位叶面积的离体材料光合效率提高几倍，能源源不断地为切口部位根源基的形成与表达提供丰富的碳源与激素能量的需求，与此还结合矿质营养液喷施补充技术为生根过程提供了所需的矿质离子养分，对于生根与壮苗起到了良好的综合效应。通过这些技术的创新与改造，实现了通常不会生根或难生根的果树品种也能快速生根的良好效果，如桃在快繁技术环境下，生根成活率可达85%-90%以上，使板栗这些极难生根品种也达80%以上，而且生根时间短，桃15-20即开始生根，板栗稍长，也只需25天就生根〔4〕。另外这种方法培育的苗木具有不定根根系特发达的特点，具有根茎比极大的特点，最适高温生长季节的移栽，做到周年快繁周年移栽的高效目的。
植物非试管快繁技术的开发运用，为果树种苗生产实现工厂化规模化快速化开辟了一个全新的空间，在加快优良品种扩繁速度，推品种结构调整进程起到了极大的作用。
植物非试管快繁技术在果树上的具体运用
植物非试管快繁技术在果树上运用最为广泛的就是各种果树的无性快繁，解决种苗繁殖中出现成活率低，周期长，难以实施标准化产业化的问题。特别是对于一些育苗周期较长的果树品种可以大大缩短种苗培育期，为生产快速捷便地生产出大量商品性一致，遗传稳定的优质苗木；另外在品种扩繁上也具有很重要的意义，对于新选育的品种如何让其以最短最快的速度达到一定的数量，满足生产所需，优化品种结构，加快更新速度具有最为快捷的效果；在无毒苗的培育上也具有特殊的效果，在人工基质及相对隔离的环境下实施，可以断绝各种土壤与昆虫传播的路径，为脱毒苗的扩繁提供了最为理想而成本最低的技术路径；在提早果树结果，实现矮化密植上也具有其它种苗不可比拟的优越性；在果园的建设上，也可以利用快繁苗进行周年移栽全年定植，打破季节的局限性；在草地果园技术的推广上，无性快繁自根苗更适于台刈修剪[5]。针对这些用途作些简要介绍，让快繁技术成为真正能让广大果农掌握与 *** 作的实用高新技术。
（1）用于果树自根苗的培养。所谓自根苗，就是发挥挖掘材料自生的生根能力，形成与植株直接联系的自生根根系。通常果树大多采用嫁接方法，利用的是砧木的根系，这是因为在传统情况下难以实现像桃梅李杏樱桃板栗杨梅枇杷芒果荔枝龙眼等枝条材料的自发根，既使有些在精细的人工管理环境下也能生根，但成活率与种苗的商品性较低，达不到产业化标准化的技术要求。而利用快繁技术可以让上述的这些品种枝段或带叶的离体材料在短期内生根，形成自根苗的不定根根系，如桃梅李杏樱桃这些核果类果树，在快繁苗床内，让一叶一芽或带叶枝段的材料在15-20天内生根，30天内就可移栽的效果；板栗杨梅枇杷芒果荔枝龙眼等难生根的果树也可达到30-45天生根移栽的效果；这样就可以大大缩短在常规下要1-2年的育苗周期，大大加快新品种的推广速度。而且上述品种采用非试管快繁后不仅对于开花结果生长特性没有影响，还具有更好的早果丰产性。这些品种在传统嫁接育苗情况下，基本上是利用本砧进行嫁接，所以采用快繁后在抗逆性上不会有变化，可以直接取枝叶进行快繁。
（2）适用于异砧类品种的快繁。对于异砧类的果树品种，如苹果梨或一些需利用砧木特殊抗性的葡萄品种，可以进行嫁接快繁法，比如苹果与梨是利用矮化砧进行矮化或者利用砧木提高抗病性与增强土壤气候适应性的品种，还有一些生长势与抗寒性较差的葡萄品种，如直接快繁失去了砧木对品种的优化驯化作用，以及特有性状的表现，如利用贝达砧木提高北方地区葡萄的抗寒冷性与一些地区的抗线虫性，利用巨峰这个长势强的品种作砧木，提高长势弱的藤稔葡萄的生长势，以实现大果大肥栽培。这些需通过砧木性状来优化品种特性的果树种类，可以采用嫁接快繁法。所谓嫁接快繁就是同时取下砧木枝段与良种枝芽，采用果树嫁接的方法进行接口绑缚结合，再把这个离体嫁接材料快繁于苗床，实现接口愈合与砧木切口生根同步进行的技术目的，这种方法虽增加了嫁接 *** 作工序，但在育苗速度还是可以达到直接快繁的效果，特别在在计算机控制的快繁苗床内，接口的愈伤会更快更好。
（3）单位面积利用率大大提高。通过上述这两种方法的结合，几乎实现了所有果树种类的快速繁殖成苗问题。育苗效率与速度大大提高，管理成本大大下降，在培育密度上通常一个平方米一批就可繁育400-1000株，以叶片不重叠为准。再加上一年可以培育至少5-6批，这样单位每平方米面积上的育苗量就几何级地超过了传统大田育苗量，可达几千株，甚至上万株，一个240平方的标准快繁苗床，年可培育果树苗木量达50-100万株，这样的高密度工厂化自动化育苗技术的实施，大大降低了生产劳动力成本，是一种当前最为有效与快速的工厂化育苗技术。
（4）采用自根苗可以提前果树的结果丰产期。在这方面的研究与探索国外起步很早，特别是台湾与日本，在桃树设施栽培及高密度栽培环境下都已开始普及与推广无性自根苗，它具有比嫁接苗更强的早果性，而且不定根的根系，更有利于树冠的控制，当前都在推广限根栽培或叫根域栽培的技术体系下，推广运用快繁无性自根苗更具技术优势，不定根的须根根系栽培后，形成的树冠更为开张，生长的枝条徒长无效枝更少，达到很好的矮化控冠之效果，在桃树李树上运用自根苗建园，前期7年中的产量大大高于嫁接苗[，其它品种也一样，都具有很强的早果性表现[6]。以色列用于这种苗进行亩栽3000-4000株的草地桃园栽培，可以使次年产量达亩产5000公斤以上，充分利用了果园早期的空间，提高了早期效益。
（7）实现脱毒苗的低成本扩繁增殖。实现脱毒苗的低成本生产，当前发达国家大力推行苹果葡萄草莓枣等脱毒苗技术，特别是苹果上运用已极为广泛，但我国生产力水平低下，培育与推广高成本的脱毒苗还存在市场与技术问题。但如果结合快繁技术实施隔离快繁，可以低成本地培育出大量脱毒苗，比土壤自然环境育苗具有更强的可 *** 作性。 *** 作上只需从已组培脱毒的母本植株上取下离体材料，再快繁于用防虫网隔绝的快繁苗床环境下进行催根育苗，就可培育出脱毒继代苗，再对继代苗进行隔离增殖培养，就是采用隔离环境下的营养液栽培，让其枝梢快速生长，再循环取枝快繁，达到脱毒苗的几何倍增效果。这种方法培育的脱毒苗成本极低，只需传统组培脱毒苗的1/10-1/20成本，便于生产推广运用，对于倡导普及脱毒苗技术具有很大的促进作用。
（8）用于草地果园建设用苗。草地果园建设时采用快繁自根苗更显技术优势，因草地果园收获后需采取特有的台刈或重截方法以控制树体的滋长，果园的郁闭；而传统的嫁接苗常会出现台刈后砧木旺长，影响树冠快速恢复与增强田间抹萌去梢的工作量，所以国外大多草地果园的建设用苗要采用没有砧木的自根苗。
一些主要果树种类的快繁方法
在计算机控制的智能化环境下，为果树枝段或带叶离体材料的发育生根创造了最佳的温光气热环境，科学有效地解决了育苗的环境问题，通过无机基质如珍珠岩的运用，解决了生根过程的透气与菌感染问题。在实施快繁果树苗木过程中环境一定的情况下，主要的技术在于材料的选择与药剂的处理，这两方面是影响生根成活率的主要因子，现把各种不同生根类型不同难易程度的果树品种所需采用的不同生根处理方法作些阐述。
（1）易生根品种的处理方法与材料要求，桃梅李杏樱桃葡萄无花果树莓等属于易生根品种，这些品种只需在带叶生长季节取材快繁都可达90%以上的生根成活率，而且根系特点发达。这类品种取材时一般取一叶一芽或一叶两芽枝段作为离体材料，运用快繁宝或吲哚丁酸低浓度浸泡作为促根处理。低浓度浸泡时间一般是100-200PPm情况下，切口浸泡1-2小时，在生产上为了提高工作效率，也可进行高浓度浸泡，即1000ppm的吲哚丁处理3-5秒钟，这些品种的快繁在枝梢旺盛生长期是生根最快成活最高的季节。
（2）较难生根品种的处理方法与材料要求，柑桔、枇杷、苹果、梨、猕猴桃等品种，要求从3-5年生以下的幼龄树上获取离体材料，最好是生长健壮无病虫害的生长枝，这些枝所制作的离体材料内源生长激素充足，叶片光合效率较高，生根容易而发达。在快繁时取带叶枝段或一叶一芽作为离体材料，切口用较高浓度的生根宝或吲哚丁酸、吲哚乙酸处理，通常浓度掌握在500-1000ppm间，进行切口浸泡处理1-2个小时。其中苹果与梨以吲哚丁酸处理效果会较好，而且可以结果滑石粉沾根处理，可以达到理想的效果。经过这些处理后，可以使大多数品种成活率达80-85%以上。
（3）极难生根的板栗杨梅芒果龙眼需进行特殊处理，这些品种因树体内含有大量阻碍生根的物质，其中单定是板栗与杨梅生根的主要抑制因子，还有芒果与龙眼材料中具有含量极高的脱落酸与其他抑根物质，对于这些品种可以先对取材的母本树进行蔗光处理，以降低体内抑根物质的合成数量，也可对于枝段的生根部位进行树上包黑纸处理，达到降低处理部位抑制物质含量，或者对所取的离体材料进行切口流水处理与硝酸银处理〔7〕，除去部份抑根物质，通过这些处理后，再行生长激素生根处理，一般配制较高浓度的吲哚丁酸浓液1000ppm，进行切口4-6小时的处理，经上述处理后，再繁于快繁苗床也可达到极高的成活率，一般可稳定在75%以上的生根率。另外，对于这些极难生根的品种一定要从幼树上取材或从已快繁成功的小苗上取材，这样可以达到更好的效果，通常经多代循环后，成活率还可大大提高。所以对于难生根品种生产上都要求形成以苗繁苗的技术体系，也就是利用已快繁成活的种苗为母本进行继代多代循环，这样可激发材料更大的生根潜能，可以使材料体内抑根物质最少化。
运用快繁技术可以使各种果树品种实现快速生根快速成苗，既使极难生根的品种也能在智能化的环境下，诱导出根源基，形成发达的根系，在生产上关键要掌握各种植物生根特性，才能有针对性地设计出快繁处理方案，实现各种果树都能达到良好的生根育苗效果。
广阔的运用前景
植物非试管快繁技术在果树上的运用是当前果树产业发展过程中涌现出来的一项全新技术，在生产科研上的运用，还有一个人们接受的过程，但从当前果苗产业的发展趋势来说，走出传统育苗制限，实现现代工厂化自动化规模化育苗是必由之路，只有这样才能强化果树产业的产前产业链，才能加快新品种的培育与扩繁推广速度，才能使广大果农以最低的成本实现品种结构调整的最优化，才能使果业发展紧跟品种换代的步伐，才能生产出更多优质的果品，以满足市场及人们生活的需要，它的运用是果树产业中的一项技术革命，具有极为广阔的发展运用前景。
Plant non- test tube quick numerous technology on fruit tree's utilization
Xu Weizhong,Zhao root,Zeng Fanqing
Agricultural Intelligentized Rapid Propagation Center of Lishui Institution of Agricultural Science
Abstract: This article comprehensive nature elaborated our country current young fruit tree production existence question and the limiting factor, have analyzed the overseas seedling production tendency, proposed the use new grows seedlings the technical ---- plant non- test tube quick numerous technology, realized the seedling production factorization, automated, the industrial production production pattern, in coor with progress of production scientific research practice concisely elaborates the quick numerous technology to apply in the fruit tree seedling cultivation various aspects with the superiority, and had pointed out each kind of different fruit tree type quick numerous technical main point, has demonstrated this technology in on the fruit tree seedling quick numerous utilization broad prospect and the market space
Key word: The plant non- test tube quick numerous technology, the fruit tree, the nursery stock, from the offspring, escapes the poisonous seedling, the factorization, Lawn orchard
作者简介：徐伟忠，研究员，浙江省丽水市农科所农业智能化快繁中心主任，2004年度全国农村青年创业致富带头人，曾主持研究开发植物非试管快繁技术，植物水生诱变技术，温室大棚控制计算机，芽苗菜智能化栽培技术等十多项技术，其中，植物非试管快繁技术经鉴定居国内领先水平，获国家星火计划项目，无形资产评估达14亿元。
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参考文献：
1、聂书海，张绪圆，李跃．荷兰果树育苗现状考察报告．河北果树1995(3)-38-38
2、周炜，曲英华．无糖组培技术在我国的研究进展．农村实用工程技术：温室园艺2005(7)-24-

Cloning in biology is the process of producing populations of genetically-identical individuals that occurs in nature when organisms such as bacteria, insects or plants reproduce asexually Cloning in biotechnology refers to processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning), or organisms More generally, the term refers to the production of multiple copies of a product such as digital media or software
Molecular cloning refers to the process of making multiple copies of a defined DNA sequence Cloning is frequently used to amplify DNA fragments containing whole genes, but it can also be used to amplify any DNA sequence such as promoters, non-coding sequences and randomly fragmented DNA It is used in a wide array of biological experiments and practical applications ranging from genetic fingerprinting to large scale protein production Occasionally, the term cloning is misleadingly used to refer to the identification of the chromosomal location of a gene associated with a particular phenotype of interest, such as in positional cloning In practice, localization of the gene to a chromosome or genomic region does not necessarily enable one to isolate or amplify the relevant genomic sequence
In practice, in order to amplify any DNA sequence in a living organism, that sequence must be linked to an origin of replication, which is a sequence of DNA capable of directing the propagation of itself and any linked sequence However, a number of other features are needed and a variety of specialised cloning vectors (small piece of DNA into which a foreign DNA fragment can be inserted) exist that allow protein expression, tagging, single stranded RNA and DNA production and a host of other manipulations
Cloning of any DNA fragment essentially involves four steps [1]
fragmentation - breaking apart a strand of DNA
ligation - gluing together pieces of DNA in a desired sequence
transfection - inserting the newly formed pieces of DNA into cells
screening/selection - selecting out the cells that were successfully transfected with the new DNA
Although these steps are invariable among cloning procedures a number of alternative routes can be selected, these are summarized as a cloning strategy’
Initially, the DNA of interest needs to be isolated to provide a DNA segment of suitable size Subsequently, a ligation procedure is used where the amplified fragment is inserted into a vector (piece of DNA) The vector (which is frequently circular) is linearised using restriction enzymes, and incubated with the fragment of interest under appropriate conditions with an enzyme called DNA ligase Following ligation the vector with the insert of interest is transfected into cells A number of alternative techniques are available, such as chemical sensitivation of cells, electroporation and biolistics Finally, the transfected cells are cultured As the aforementioned procedures are of particularly low efficiency, there is a need to identify the cells that have been successfully transfected with the vector construct containing the desired insertion sequence in the required orientation Modern cloning vectors include selectable antibiotic resistance markers, which allow only cells in which the vector has been transfected, to grow Additionally, the cloning vectors may contain colour selection markers which provide blue/white screening (α-factor complementation) on X-gal medium Nevertheless, these selection steps do not absolutely guarantee that the DNA insert is present in the cells obtained Further investigation of the resulting colonies is required to confirm that cloning was successful This may be accomplished by means of PCR, restriction fragment analysis and/or DNA sequencing
Cloning a cell means to derive a population of cells from a single cell In the case of unicellular organisms such as bacteria and yeast, this process is remarkably simple and essentially only requires the inoculation of the appropriate medium However, in the case of cell cultures from multi-cellular organisms, cell cloning is an arduous task as these cells will not readily grow in standard media
A useful tissue culture technique used to clone distinct lineages of cell lines involves the use of cloning rings (cylinders)[2] According to this technique, a single-cell suspension of cells which have been exposed to a mutagenic agent or drug used to drive selection is plated at high dilution to create isolated colonies; each arising from a single and potentially clonally distinct cell At an early growth stage when colonies consist of only a few of cells, sterile polystyrene rings (cloning rings), which have been dipped in grease are placed over an individual colony and a small amount of trypsin is added Cloned cells are collected from inside the ring and transferred to a new vessel for further growth
Cloning in stem cell research
Main article: Somatic cell nuclear transfer
Somatic cell nuclear transfer can also be used to create a clonal embryo The most likely purpose for this is to produce embryos for use in research, particularly stem cell research This process is also called "research cloning" or "therapeutic cloning" The goal is not to create cloned human beings, but rather to harvest stem cells that can be used to study human development and to potentially treat disease While a clonal human blastocyst has been created, stem cell lines are yet to be isolated from a clonal source[3]
Horticultural
The term clone is used in horticulture to mean all descendants of a single plant, produced by vegetative reproduction or apomixis Many horticultural plant cultivars are clones, having been derived from a single individual, multiplied by some process other than sexual reproduction As an example, some European cultivars of grapes represent clones that have been propagated for over two millennia Other examples are potato and banana Grafting can be regarded as cloning, since all the shoots and branches coming from the graft are genetically a clone of a single individual, but this particular kind of cloning has not come under ethical scrutiny and is generally treated as an entirely different kind of operation
Many trees, shrubs, vines, ferns and other herbaceous perennials form clonal colonies Parts of a large clonal colony often become detached from the parent, termed fragmentation, to form separate individuals Some plants also form seeds asexually, termed apomixis, eg dandelion
Parthenogenesis
Clonal derivation exists in nature in some animal species and is referred to as parthenogenesis (reproduction of an organism by itself without a mate) An example is the "Little Fire Ant" (Wasmannia auropunctata), which is native to Central and South America but has spread throughout many tropical environments
Reproductive cloning
Reproductive cloning uses "somatic cell nuclear transfer" (SCNT) to create animals that are genetically identical This process entails the transfer of a nucleus from a donor adult cell (somatic cell) to an egg which has no nucleus If the egg begins to divide normally it is transferred into the uterus of the surrogate mother
Such clones are not strictly identical since the somatic cells may contain mutations in their nuclear DNA Additionally, the mitochondria in the cytoplasm also contains DNA and during SCNT this DNA is wholly from the donor egg, thus the mitochondrial genome is not the same as that of the nucleus donor cell from which it was produced This may have important implications for cross-species nuclear transfer in which nuclear-mitochondrial incompatibilities may lead to death
Dolly the Sheep
Main article: Dolly the Sheep

Dolly (1996-07-05 – 2003-02-14), a Finn Dorsett ewe, was the first mammal to have been successfully cloned from an adult cell, though the first actual thing to be cloned, was a tadpole in 1952[1] She was cloned at the Roslin Institute in Scotland and lived there until her death when she was six On 2003-04-09 her stuffed remains were placed at Edinburgh's Royal Museum, part of the National Museums of Scotland
Dolly was publicly significant because the effort showed that the genetic material from a specific adult cell, programmed to express only a distinct subset of its genes, could be reprogrammed to grow an entire new organism Before this demonstration, there was no proof for the widely spread hypothesis that differentiated animal cells can give rise to entire new organisms
Cloning Dolly the sheep had a low success rate per fertilized egg; she was born after 277 eggs were used to create 29 embryos, which only produced three lambs at birth, only one of which lived Seventy calves have been created from 9,000 attempts and one third of them died young; Prometea took 328 attempts Notably, although the first clones were frogs, no adult cloned frog has yet been produced from a somatic adult nucleus donor cell
There were early claims that Dolly the Sheep had pathologies resembling accelerated aging Scientists speculated that Dolly's death in 2003 was related to the shortening of telomeres, DNA-protein complexes that protect the end of linear chromosomes However, other researchers, including Ian Wilmut who led the team that successfully cloned Dolly, argue that Dolly's early death due to respiratory infection was unrelated to deficiencies with the cloning process
Species cloned
Further information: List of animals that have been cloned
The modern cloning techniques involving nuclear transfer have been successfully performed on several species Landmark experiments[clarify] in chronological order:
Tadpole: (1952) Many scientists questioned whether cloning had actually occurred and unpublished experiments by other labs were not able to reproduce the reported results[citation needed]
Carp: (1963) In China, embryologist Tong Dizhou cloned a fish He published the findings in a Chinese science journal which was never translated into English[4]
Mice: (1986) was the first successfully cloned mammal; Soviet scientists Chaylakhyan, Veprencev, Sviridova, Nikitin had mice "Masha" cloned Research was published in the magazine "Biofizika" volume ХХХII, issue 5 of 1987[clarify][5]
Sheep: (1996) From early embryonic cells by Steen Willadsen Megan and Morag[citation needed] cloned from differentiated embryonic cells in June 1995 and Dolly the sheep from a somatic cell in 1997[6]
Human: (November 1998) hybrid embryo created from leg cells and a cleaned cow egg - not allowed to implant in a womb, nor develop, nor be born due to ethical issues[citation needed]
Rhesus Monkey: Tetra (female, January 2000) from embryo splitting[7][clarify]
Gaur: (2001) was the first endangered species cloned[8]
Cattle: Alpha and Beta (males, 2001) and (2005) Brazil[9]
Cat: CopyCat "CC" (female, late 2001), Little Nicky, 2004, was the first cat cloned for commercial reasons[citation needed]
Mule: Idaho Gem, a john mule born 4 May 2003, was the first horse-family clone[citation needed]
Horse: Prometea, a Haflinger female born 28 May 2003, was the first horse clone[citation needed]
Human cloning
Main article: Human cloning
Human cloning is the creation of a genetically identical copy of an existing or previously existing human The term is generally used to refer to artificial human cloning; human clones in the form of identical twins are commonplace, with their cloning occurring during the natural process of reproduction There are two commonly discussed types of human cloning: therapeutic cloning and reproductive cloning Therapeutic cloning involves cloning cells from an adult for use in medicine and is an active area of research: while reproductive cloning would involve making cloned human beings Such reproductive cloning has not been performed and is illegal in many countries A third type of cloning called replacement cloning is a theoretical possibility, and would be a combination of therapeutic and reproductive cloning Replacement cloning would entail the replacement of an extensively damaged, failed, or failing body through cloning followed by whole or partial brain transplant
The various forms of human cloning are controversial[10] There have been numerous demands for all progress in the human cloning field to be halted Some people and groups oppose therapeutic cloning, but most scientific, governmental and religious organizations oppose reproductive cloning The American Association for the Advancement of Science (AAAS) and other scientific organizations have made public statements suggesting that human reproductive cloning be banned until safety issues are resolved [11] Serious ethical concerns have been raised by the idea that it might be possible in the future to harvest organs from clones Some people have considered the idea of growing organs separately from a human organism - in doing this, a new organ supply could be established without the moral implications of harvesting them from humans Research is also being done on the idea of growing organs that are biologically acceptable to the human body inside of other organisms, such as pigs or cows, then transplanting them to humans, a form of xenotransplantation
The first human hybrid human clone was created in November 1998, by American Cell Technologies[12] It was created from a man's leg cell, and a cow's egg whose DNA was removed It was destroyed after 12 days Since a normal embryo implants at 14 days, Dr Robert Lanza, ACT's director of tissue engineering, told the Daily Mail newspaper that the embryo could not be seen as a person before 14 days While making an embryo, which may have resulted in complete human had it been allowed to come to term, according to ACT: "[ACT's] aim was 'therapeutic cloning' not 'reproductive cloning'"
On January, 2008, Wood and Andrew French, Stemagen's chief scientific officer in California, announced that they successfully created the first 5 mature human embryos using DNA from adult skin cells, aiming to provide a source of viable embryonic stem cells Dr Samuel Wood and a colleague donated skin cells, and DNA from those cells was transferred to human eggs It is not clear if the embryos produced would have been capable of further development, but Dr Wood stated that if that were possible, using the technology for reproductive cloning would be both unethical and illegal The 5 cloned embryos, created in Stemagen Corporation lab, in La Jolla, were destroyed[13]
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