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研究生:楊金昌
研究生(外文):Yong, Kim-Chong
論文名稱:數種甘藷屬植物生育特性及其葉肉原生質體之研究
論文名稱(外文):Studies on Plant Growth Characteristics and Leaf Protoplasts of Several Ipomoea Species
指導教授:賴光隆賴光隆引用關係
指導教授(外文):Kwan-Long Lai
學位類別:博士
校院名稱:國立臺灣大學
系所名稱:農藝學系
學門:農業科學學門
學類:一般農業學類
論文種類:學術論文
論文出版年:1997
畢業學年度:85
語文別:中文
論文頁數:124
中文關鍵詞:甘藷甘藷近緣野生種原生質體電融合
外文關鍵詞:sweet potatoIpomoea wild speciesprotoplastelectrofusion
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甘藷(Ipomoea batatas (L.) Lam.)是本省重要的根類作物,在亞洲的栽
培面積及產量為世界之冠,是很值得研究推廣的作物。野生種源的收集研
究利用在甘藷改良、基礎研究及育種上亦極為重要。本研究的材料為三種
甘藷屬植物,包括栽培種甘藷台農66號、近緣野生種甘藷 I. triloba及
I. trifida 〔具二倍體(2n=2x)、四倍體(2n=4x)與六倍體(2n=6x)〕,建
立基本植株形態資料、葉肉原生質體分離、培養系統,並探討甘藷屬原生
質體電融合的可行性。首先建立參試材料試管苗,參試材料種子經滅菌後
播種於含0.7﹪洋菜、3﹪蔗糖、pH 5.7之MS培養基上,可順利萌芽長成植
株,經過含不同濃度NAA之MS培養基繼代培養試驗,找出適合的繼代培養
基。結果顯示帶一至二個腋芽的I. triloba莖段繼代培養於含0.1 mg/l
NAA的MS固態培養基較佳,而I. trifida 2x、4x及6x則分別繼代培養於含
0.1 mg/l、0.3 mg/l及0 mg/l NAA之MS固態培養基較佳。將栽培種甘藷台
農66號及三種I. trifida材料自試管移出種植於三種不同日/夜溫度的自
然光照玻璃溫室中,調查植株形態及生長習性。結果顯示三種I. trifida
材料之植株形態及生長習性上如多年生、自交不和合性及根中含澱粉粒等
特性,皆與栽培種甘藷台農66號相似。栽培種甘藷台農66號及三種I.
trifida之葉片長寬、葉柄長度與直徑、莖直徑及第五節間長度均隨栽培
之日/夜溫度升高而有增加的趨勢。葉形的變化、葉片表皮細胞、莖的粗
細則隨染色體倍數性增加而增加。在原生質體分離培養試驗方面,栽培種
甘藷台農66號之葉肉在酵素液處理前,先經浸泡甘露醣醇或山梨醣醇溶液
處理,確可增加原生質體的分離量。三種I. trifida材料葉肉於含有4﹪
cellulase RS 、0.5﹪ pectolyase Y-23、0.5﹪ hemicellulase、0.4M
甘露醣醇、1/4MS無機鹽類、1 mg/l thiamine-HCl、1 mg/l pyridoxine-
HCl 、10 mg/l nicotinic acid及100 mg/l myo-inositol的酵素液中可
順利分離得到原生質體,I. trifida 2x、4x及6x每公克新鮮葉片分別可
分離得到1.45x107±0.32x107、3.95x106±1.22x106、3.5x106±1.65
x106 個原生質體,培養於含0.1 mg/l 2,4-D及0.5 mg/l KIN之1/2修正MS
液態培養基可分裂形成細胞團,經平埋培養及移至固態培養基可形成癒合
組織。電融合條件試驗方面,顯示在交流電場及直流電場分別為600 V/cm
及1800 V/cm時有較佳的結果。融合原生質體均可培養形成癒合組織,其
中自栽培種甘藷台農66號及I. triloba融合原生質體誘導之癒合組織在含
NAA及BA的分化培養基上可分化器官形成根,但經分析其過氧化氫同功酵
素電泳圖譜及細胞染色體均表現與栽培種甘藷台農66號相類似。而誘導自
I. trifida 2x及4x融合原生質體之癒合組織則有呈現異於兩融合親本間
的過氧化氫同功酵素電泳圖譜及其染色體數2n等於90或多於90之情形。
Sweet potato (Ipomoea batatas (L.) Lam.) is one of the most
important root crops in Taiwan. It''s area harvested and total
production in Asia is top of the world. Since sweet potato is a
crop which is always propagated by asexual propagation method ,
it would be appropriate to carry out those update research
methods. In this research first of all the plant characteristics
of sweet potato cultivar Tainung No.66 (TNG66), two closely
related wild species of sweet potato I. triloba and I. trifida
〔containing diploid (2n=2x),tetraploid(2n=4x) and hexaploid(2
n=6x)〕were investigated, then established the methods of
isolation and culture of their leaf mesophyll protoplast. The
condition of electrofusion between their protoplasts and culture
were also studied. The characteristics of fusant were examined
with isozyme zymograms and chromosome numbers respectively.Seeds
of closely related wild species were firstly disinfected and
sown on the aseptic solidified MS medium containing 3% sucrose
and 0.7% agar-agar with pH5.7. After the plant were established
in vitro, their stem segment which contained one to two axillary
buds were subcultured to fresh medium containing different
concentration of NAA. The results showed that I. triloba , I.
trifida 2x ,4x and 6x grew better in medium containing 0.1 mg/l,
0.1 mg/l, 0.3 mg/l and 0 mg/l NAA respectively.Plants of sweet
potato cultivar TNG66 and three different ploidy plants of I.
trifida were transplanted to natural light phytotrone with
different day/night temperatures. Plants characteristics and
growth habitat were investigated, there are several plant
characteristics and growth habitat such as self-incompatibility,
starch containing in roots and perennial growth, which are
similar in cultivar TNG66 and three different ploidy plants of
I. trifida. Width and length of leaf ,length and diameter of
petiole, diameter of stem and length of 5 th internode of
cultivar TNG66 and those I. trifida increased when the growing
day/night temperature were increased. Variation of leaf shape,
periderm cell on leaf and thickness of stem were increased with
the increment of ploidy among those I. trifida. Protoplast
isolation of leaf mesophyll of sweet potato cultivar TNG66 were
found to be improved greatly if the leaf mesophyll were
presoaked in mannitol or sorbitol solution before enzyme
solution treatment. Leaf mesophyll protoplasts of the three
different ploidy plants of I. trifida could be isolated
successfully by treatment of enzyme solution containing 4﹪
cellulase RS, 0.5﹪pectolyase Y-23, 0.5﹪hemicellulase, 0.4 M
mannitol, 1/4 MS inorganic salts, 1 mg/l thiamine-HCl, 1 mg/l
pyridoxine-HCl , 10 mg/l nicotinic acid及100 mg/l myo-inositol
without pretreatment. Colonies were induced respectively from
the three ploidy I. trifida protoplasts when they were cultured
in MS liquid medium with 0.1 mg/l 2,4-D and 0.5 mg/l KIN.
Protoplasts yield per gram leaf fresh weight of I. trifida 2x,4x
and 6x were 1.45x107±0.32x107, 3.95x106±1.22x106 and 3.5x106
±1.65x106 respectively, and callus were formed after the
colonies were plated and transfered to solid medium.The
electrofield strength of protoplast electrofusion condition were
studied. The result showed that adequate ac-field and dc-field
for protoplast inducing electrofusion of sweet potato cultivar
TNG66 and I. triloba were 600 V/cm and 1800 V/cm. Callus could
be induced from the electrofused protoplasts of sweet potato
cultivar TNG66 and I. triloba after cultured. Root organs were
regenerated when they were transferred to 1/2 modified MS medium
containing NAA and BA. The peroxidase isozyme zymogram and
chromosome number of those callus similar to that of sweet
potato cultivar TNG66. Callus induced from electrofused
protoplasts of I. trifida 2x and 4x did show different zymogram
of peroxidase isozyme from both protoplast donors. Their
chromosome number showed 2n=90 or more.
封面
目錄
頁次
表目錄
圖目錄
中文摘要
英文摘要
縮寫字及全名對照
第一章 緒言
第二章 前人研究
第三章 供試植物之生育特性
第四章 甘藷屬葉肉原生質體分離與培養特性
第五章 甘藷屬葉肉原生質體之誘融與培養條件之探討
第六章 誘融原生質體過氧化氫同功酵素及染色體檢定
第七章 綜合討論
參考文獻
附表1
附表2
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