About 35 years ago, having cracked how I could make a lot of daffodil shoots in test tubes (that’s another story!), I was faced with a great hurdle that for the life of me I had no idea how to tackle. How could I turn these shoot clumps into little bulbs for planting in the field? After all, my PhD’s goal was to produce a complete protocol from multiplying massive number of daffodils/narcissus shoot clumps in test-tubes and getting these growing ‘normally’ in the field. Based on literature reviews, I knew that plating shoot clumps had been successfully done by other researchers. But this had two major disadvantages:
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- There would be a danger of these shoot clumps not acclimatizing well when planted out in the glass house. 25 – 30% casualties would be “normal”.
- These shoot clumps were behaving like young narcissus seedlings and would need up to five years to grow to flowering stage.
My late supervisor, Dr. Barbara M.R. Harvey suggested I should look at other plant models in tissue culture for inspirations. The light-bulb moment came when I had a chat with Dr. Nikki Evans on how she got her potato shoots in test-tubes to form small tubers. Evans used high sugar content in her culture medium and I went along this line. A few months later, I was elated to see little bulbs (which I termed ‘bulbils”) formed in my test-tubes given high dose of sucrose in their culture media. That was the turning point of my PhD and 6 months later I submitted my thesis…and the rest as they say “was history”!
I owed it to my two supervisors, Dr. Barbara Harvey and Dr. Christopher Selby for putting their feet down shortly after I had completed the “bulbils” experiment by stopping me from doing any more work in the lab (they literally banished me from my lab!) and by pushing me to write my doctorate thesis which I completed about 6 months later! This was the most important turning point! My research project goals were attained, but my doctoral studies goal was still not reached. I had to write, submit and defend my thesis (successfully) to earn my PhD!
A few months after that (in December 1990), I was conferred the degree of Doctor of Philosophy by my alma mater, the Queen’s University of Belfast. Shortly after, I left Belfast to take up a post-doctoral research officer post at the National University of Singapore.
It was around late 1991 that I received the great news from Dr. Harvey, she wrote in an email: “Chow, some of the “bulbils” that you had planted out in mid 1990 are flowering!”
This was exciting news indeed because:
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- it proofed that my bulbils, after going through the “stressful” process of my protocol were not any different from those multiplied conventionally;
- of more significance is the fact that this meant my protocol had cut the “shoot clumps/seedlings to flowering bulb stage”by at least 3 – 4 years!
I wished I had the chance and resources to carry on with this work to collect more data on this observation but disappointingly this was not the case.
In research work, you do need to have a keen eye to spot trends & changes but what you need most is the guidance of experienced researchers. I was lucky to have both! The keen eye helped me to spot the resemblance between the potato tuberization process and the bulb formation of narcissus/daffodils. The guidance of my supervisors ensured that I scaled the last but most crucial turning point of my PhD journey. They put a stop to my laboratory work and made sure that I stuck to the PhD research schedule (and more importantly, my scholarship tenure) to compose and submit my PhD thesis way before the last of my scholarship cheques was issued!
If you are interested, a brief introduction to a paper published by my supervisors and I have just been written & published in Kudos by me.
中文版：
大约35年前，我成功的在试管里培养出了很多水仙花的嫩芽(其实这是首个转折点，也是另一个故事！)，我面临着一个巨大的障碍，费尽了心思我根本不知道该如何去解决。我怎么能把这些芽丛变成小鳞茎以便他们可以顺利的在田间栽种呢？毕竟，我博士科研的目标是产生一个完整的方案，通过组织培养大量的生产水仙花芽丛，并使这他们可以在田间“正常”生长。根据文献回顾，我知道其他研究人员已经成功地将组织培养的水仙芽丛栽种到田间。但栽种这组培水仙芽丛有两个主要缺点：
● 将这些芽丛从试管移植到温室中，有它们不能很好地适应环境的危险。25-30%的损失将是“正常的”。
● 这些芽丛的本质就像刚发芽的水仙幼苗，将需要大概长达5年时间从成长到开花阶段。
我已故的导师芭芭拉·哈维博士建议我从其他的组织培养植物模型中寻找灵感。当我和尼基·埃文斯博士聊到她是如何在试管中培育出小块茎的时候，我灵光一闪。埃文斯博士在她组织培养土豆实验中使用了高含糖量的培养基就使得小土豆在试管内形成，我就跟随她的实验方向这样做。几个月后，我欣喜地看到了我实验试管里有些芽丛已经有所变化：可以观察到在我给予高剂量的蔗糖培养基的水仙芽丛里有小鳞茎（我称之为“珠芽”）的形成。那就是我读博士的另一个转折点。6个月后，我成功的提交了我的博士论文…其余的，就是英语术语说的”是历史”！
在我的这一部分工作中，我要感谢埃文斯博士慷慨地分享她的想法。
我的两位博士导师，芭芭拉·哈维博士和克里斯托弗·塞尔比博士在我完成了“珠芽”实验后不久就很精明的阻止了我在实验室里做更多的实验(他们确实是把我“赶出“了实验室！) 。通过他们的催促，我全心全力的写好我的博士论文，大约6个月后我完成了这篇论文！这是最重要的转折点！因为当时我的研究项目目标已经实现，但我的博士研究目标仍未达到。为了获得博士学位，我必须写作、提交和成功的答辩我的论文！
几个月后（1990年12月），我被母校贝尔法斯特女王大学授予哲学博士学位。不久之后，我离开贝尔法斯特，到新加坡国立大学担任博士后研究员。
大约在1991年底，我收到了哈维博士的好消息，她在一封电子邮件中写道：“小周，你在1990年中所种下的一些‘珠芽’正在开花了！”
这确实是一个令人兴奋的消息，因为：
● 它证明了，在经历了我组织培养环境的“压力”及过程之后，我的珠芽和那些按惯例所被繁殖的鳞茎是没有什么不同；
● 更重要的是，这意味着我的方式方法成功的将“芽丛/幼苗到开花鳞茎期”缩短了至少3到4年！
当时我希望我有机会和资源在这方面进行更深入的研究工作，收集更多的数据，但令人失望的是，这已经是不可能的。
在研究工作中，你确实需要洞察力去敏锐地观察趋势和变化，但你最需要的是有经验研究人员的指导。我很幸运两者都有！敏锐的眼睛帮助我发现了土豆块茎化过程和水仙花鳞茎形成之间的相似之处。在导师的指导下，我攀登了博士之旅的最后一个但也是最关键的转折点。我的两位导师在我完成了“珠芽实验”后就停止了我的实验室工作，并确保我跟随学校设定博士研究时间表(更重要的是，我的奖学金任期)，在我的最后一份奖学金支票被发出之前完成撰写和提交我的博士论文！
如果你有兴趣，我刚刚在Kudos杂志上发表了一篇文章是描述我两位导师和本人发表的一篇有关水仙芽丛形成珠芽的论文。