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1、专业好文档关于植物组织培养的实习报告植物的组织培养是以植物生理学为基础，近几十年发展起来的一项无性繁殖的新技术。广义上又叫离体培养，指将植物的离体器官、组织、细胞以及去除细胞壁的原生质体，通过无菌操作，在人工控制环境下，培养在人工配制的培养基上对其进行克隆，使其生长、分化并成长为完整植株的过程。本人于2010年11月17日至2011年1月20日在北京太阳宫园林绿化有限公司牛栏山基地自行实习。我的实习内容主要包括MS培养基的制备，萱草、玉簪的增殖培养，草莓茎尖的诱导。具体实践过程如下：一、MS培养基的制备1.浓缩贮备液 需配制一系列的浓缩贮备液为大量元素（浓缩20x）、微量元素（浓缩1000x）
2、、铁盐（浓缩100x）、除蔗糖之外的有机物质（浓缩1000x）。所有的贮备液都应贮存于塑料瓶中或玻璃瓶中，置冰箱中保存，其中铁盐贮备液必须贮存于琥珀色玻璃瓶中。2.制备1LMS空白培养基 先量取贮备液50ml、贮备液1ml、贮备液10ml、贮备液1ml至量筒中加适量蒸馏水定容至1L，并倒入容器中加热；然后称取蔗糖30g/L，琼脂4.5g/L搅拌至完全溶解；再用NaOH和HCl调至 pH值为5.8，分装至玻璃瓶中（30ml/瓶），最后放入高压锅中灭菌，在压力为108kPa、温度121下灭菌20min即可。二、萱草的增殖培养将增殖培养出的丛生芽与愈伤组织一起，分割转接到MS+BA2.0mg/L+I
3、BA0.1mg/L的培养基上进行培养。培养室温度为252，光照强度约12001500LX，光照时间为10小时。经过2030天芽与愈伤的不断增殖生长，即可分化出丛生芽。每隔3045天，将分化的丛生芽切成23株再接种在增殖培养基上，每次可扩大5倍。三、玉簪的增殖培养将增殖分化出的丛生芽接种到MS+BA2.0mg/L+KT2.0mg/L+NAA0.05 mg/L培养基上进行增殖培养，培养室温度为252，光照强度约15002000LX，光照时间为12小时。每隔3045天将分化出丛生芽切成单株再接种到增殖培养基上，每次可扩大45倍（淘汰增殖少而慢的材料）。增殖繁殖代数绿叶品种控制在24代以内，花叶品种控
4、制在16代以内。四、草莓茎尖的诱导（一）外植体的选择与消毒1.外植体的选择 草莓的组织培养以茎尖作为外植体效果较好，常用于草莓快繁脱毒技术。摘取基地中草莓红颜品种新抽出的健壮无病的匍匐茎45cm长作为外植体材料，进行丛生芽的诱导。2.外植体的消毒 用自来水冲洗一小时，洗刷表面污物及绒毛，然后加入84消毒液与洗涤灵清洗10min，同时利用离心力转圈摇动瓶子使外植体表面的污染物被析出，最后用自来水冲洗数次，直到没沫为止将水倒掉。移置于超净工作台上，将冲洗后材料再截成23cm长，用75%酒精浸泡1520s，用无菌水冲洗1次，然后用01的升汞(HgCl2)溶液浸泡杀菌510min，再用无菌水冲洗56次
5、，少量多次，最后用无菌滤纸吸去外植体表面的水分，待接种。（二）接种与培养1.接种 在无菌条件下，将已经过消毒的匍匐茎外植体，用解剖刀剥取其微茎尖，接种到MS基本培养基中。刚接种的外植体需进行暗培养（3天）或是保持较低温度等措施，均可减轻材料褐化的发生；增加光照或延长光照时间都可降低材料玻璃化的产生。2.培养 建立无菌体系得到无菌苗后，接种到MS+BA0.5mg/L+IBA0.1mg/L芽诱导培养基上培养，培养室温度为252，光照强度约20003000LX，光照时间为12小时。20d左右无菌苗增厚，切口有瘤状突起，即愈伤组织，继续培养即可由它分化出不定芽。将不定芽与愈伤组织一起，分割转接到MS+
6、BA1.0mg/L+NAA0.05mg/L增殖培养基上培养，经过1520天芽与愈伤的不断增殖生长，即可分化出丛生芽。每隔2030天将分化的丛生芽转接至增殖培养基上，每次可扩大810倍。五、实习中发现的问题我是从2010年6月份开始深层次接触植物组织培养这个行业的，在学校时虽然学习过组培的相关知识，但只是理论知识，实际操作方面接触并不多。这次的实习刚好是我在逐步熟悉组培这个行业的过程中进行的，使我更加深刻的发现了自身存在的一些问题，具体如下：1.增强专业知识，学以致用 初入组培这个行业，经验不足，书本上的理论知识仍需与实际相结合。在专业知识上与自己本职工作需求还存在着一定的差距，需要不断增强自己
7、的专业性知识，学以致用，并以书中所写为依据，根据实际情况适时变通。2.熟练操作技术，提高速度 初接触时，由于操作技术不熟练，导致组培材料大量被污染，损失严重。因此在操作过程中越规范材料被污染的可能性就越小，再在此基础上提高速度，适应工厂化生产。3.明确问题的实质与根本 在工作中缺乏新思维，对事物发展中存在的问题认识不够准确，把握不住发展的必然规律。对实习中遇到的困难和矛盾，有时把握不住主要矛盾和矛盾的主要方面，抓不住问题的实质和根本。4.明确有效的目标或计划 像是周计划、月计划、学期计划等等。没有目标与计划就像是不知道方向的船，在海洋里任意漂游，最终会在海洋中消失。为自己制定明确的目标与计划，
8、一步一步认真的按计划进行，实现目标。六、总结初学植物组织培养时，对于一切都很陌生的我感到了困惑，不知该从哪里入手，虽然之前也有看过相关的书籍，但毕竟与实际还是有差异的。这次的实习经历让我更加充分、更加深入的了解到植物组织培养的相关知识。每次看到经过自己动手接种、增殖后慢慢长大的组培苗，心里都会有小小的成就感。通过这次综合性的实习，使我掌握了现代生物技术手段与运用能力，训练了实际操作技能。将我所学的基础理论、基础知识和基本技能运用于实践之中，将所学的知识转化为能力，培养实际工作能力、科研能力。这次的实习对于我来说受益匪浅，是不断学习、不断充实自己的一次机会，也是积极探索、逐步成长的一次经历，我会
9、在不断增强自己专业知识的同时，利用现有的优越条件，理论与实际相结合，做到全面性发展。Winger Tuivasa-Sheck, who scored two tries in the Kiwis 20-18 semi-final win over England, has been passed fit after a lower-leg injury, while Slater has been named at full-back but is still recovering from a knee injury aggravated against USA.Both sides boa
10、st 100% records heading into the encounter but Australia have not conceded a try since Josh Charnleys effort in their first pool match against England on the opening day.Aussie winger Jarryd Hayne is the competitions top try scorer with nine, closely followed by Tuivasa-Sheck with eight.But it is re
11、cently named Rugby League International Federation player of the year Sonny Bill Williams who has attracted the most interest in the tournament so far.The Kiwi - with a tournament high 17 offloads - has the chance of becoming the first player to win the World Cup in both rugby league and rugby union
12、 after triumphing with the All Blacks in 2011.Id give every award back in a heartbeat just to get across the line this weekend, said Williams.The (lack of) air up there Watch mCayman Islands-based Webb, the head of Fifas anti-racism taskforce, is in London for the Football Associations 150th anniver
13、sary celebrations and will attend Citys Premier League match at Chelsea on Sunday.I am going to be at the match tomorrow and I have asked to meet Yaya Toure, he told BBC Sport.For me its about how he felt and I would like to speak to him first to find out what his experience was.Uefa hasopened disci
14、plinary proceedings against CSKAfor the racist behaviour of their fans duringCitys 2-1 win.Michel Platini, president of European footballs governing body, has also ordered an immediate investigation into the referees actions.CSKA said they were surprised and disappointed by Toures complaint. In a st
15、atement the Russian side added: We found no racist insults from fans of CSKA. Baumgartner the disappointing news: Mission aborted.The supersonic descent could happen as early as Sunda.The weather plays an important role in this mission. Starting at the ground, conditions have to be very calm - winds
16、 less than 2 mph, with no precipitation or humidity and limited cloud cover. The balloon, with capsule attached, will move through the lower level of the atmosphere (the troposphere) where our day-to-day weather lives. It will climb higher than the tip of Mount Everest (5.5 miles/8.85 kilometers), d
17、rifting even higher than the cruising altitude of commercial airliners (5.6 miles/9.17 kilometers) and into the stratosphere. As he crosses the boundary layer (called the tropopause),e can expect a lot of turbulence.The balloon will slowly drift to the edge of space at 120,000 feet ( Then, I would a
18、ssume, he will slowly step out onto something resembling an Olympic diving platform.They blew it in 2008 when they got caught cold in the final and they will not make the same mistake against the Kiwis in Manchester.Five years ago they cruised through to the final and so far history has repeated its
19、elf here - the last try they conceded was scored by Englands Josh Charnley in the opening game of the tournament.That could be classed as a weakness, a team under-cooked - but I have been impressed by the Kangaroos focus in their games since then.They have been concentrating on the sort of stuff tha
20、t wins you tough, even contests - strong defence, especially on their own goal-line, completing sets and a good kick-chase. Theyve been great at all the unglamorous stuff that often goes unnoticed in the stands but not by your team-mates.It is as though their entire tournament has been preparation f
21、or the final.In Johnathan Thurston, Cooper Cronk, Cameron Smith and either Billy Slater or Greg Inglis at full-back they have a spine that is unmatched in rugby league. They have played in so many high-pressure games - a priceless asset going into Saturday.The Kiwis are a lot less experienced but winning a dramatic match like their semi-final against England will do wonders for their confidence.They defeated Australia in the Four Nations final in 2010 and the last World Cup, and know they can rise to the big occasion.5
