无核葡萄及其杂种胚败育与移栽的某些生理生化变化的研究
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摘要
本文通过无核葡萄(无核白自交、无核白×爱莫无核),有核葡萄(北醇、泰山 1
号、泰山 12 号)以及无核葡萄与中国野生葡萄杂种(红光无核×山葡萄)的胚珠在不
同时间离体培养,并结合生理生化指标测定发现:不同的无核葡萄及其杂种胚败育的时
间不同,胚败育时,SOD、POD、CAT 酶活性下降、膜脂过氧化(MDA)水平上升,可溶性
糖与可溶性蛋白质降低,可溶性淀粉变化不大,进行胚挽救时,应根据不同的杂交组合,
在最佳时间即胚发育程度最高,SOD、POD、CAT 三种保护酶活性最高,MDA 水平最低时
取胚珠培养,效果较好。
对胚挽救获得的试管苗经不同时间的炼苗,测定叶片气孔,叶绿素含量及保护酶
(SOD)活性,发现随闭瓶开瓶炼苗时间的延长,气孔开张度逐渐降低,关闭指数增加,
叶绿素含量增加,保护酶(SOD)活性增加。这充分说明了分步炼苗的必要性,经过分
步炼苗,苗叶生长健壮,保护组织形成,气孔恢复自我调节功能,根系吸收能力增强,
从而提高了试管苗对外界环境条件的适应能力,有利于移栽成活率的进一步提高。并提
出了无核葡萄及其杂种胚挽救试管苗最佳配套移栽技术:
①培育优质健壮试管生根苗;
②移栽基质和苗根部消毒;
③ 蛭石或珍珠岩(上层)与 田间土(下层)分层移栽,下面土层提供根系生长
2 3
5 5
发育的营养,而上面沙层提高根基部透气性,可防止根茎部霉烂;
④温室自然光闭口炼苗 7d,过渡到开口炼苗 3～7d;
⑤移栽初期注意保湿和适度遮阳,培养室条件下培养。
同时移栽后加强病虫防治和水肥管理,在自然条件下生长,最后定植到田间。
Different seedless grapes and their hybrids had different embryo
abortive period. During embryo abortion,the activities of SOD, POD,
CAT decreased,the level of MDA the soluble protein and soluble sugar
increased and the change of soluble starch was small.So during embryo
rescue process,we should select the optimum time to culture ovules
according to different cross combinations,that is the embryo
development level was best,the activities of SOD, POD, CAT is highest
and the level of MDA is lowest.All these results were obtained through
culturing ovules in vitro of seedless grapes (self-pollinated
Sultanina,Sultanina×Olmo),seededgrapes (Beichun, Taishan-1,
Taishan-12) and pollinations between seedless grapes and Chinese
wild grapes (Flame Seedless×Heilongjiang Seedling) and destmining
the physiological and biochemical index.
The open degress and the index of leaf stomates,the content
of chlorophyll and the activities of protective enzyme (SOD) were
determined.After tissue culture plants obtained from embryo rescue
being domesticated through different time.The results showed: the
prolonging domesticatng time by closed-unclosed bottle,the open
degress of stomate dropped,the index of stomate close,the contents
of chlorophyll and the activities of protective enzymes (SOD)
incressed. These indicated that it is necessary to domesticate plants
step by step.And by this kind of domestication,the plants growed
stronghy,the protective tissue developed well,self-adjusting
function of stomate recovered and the absorption capacity of roots
increased. So the adaptability of the embryo rescue plants to
envierment is improved,thus it is benefit for transplanting survival
rate.In this study,the optimam transplanting technology of seedless
grapes and their hybrids was obtained:
First,good quality tissue culture plants with roots were
necessary.
Second,the transplanting matrix and the roots of plants must be
steriled.
Third,the optimam matrix is vermiculite or pearlit (upper
2
5
layer) and 3 nutrient soil (under layer) the nourishing soil can
5
provide the nourishment for root growing. While the vermiculite or
pearlite can improve capacity.Thus it can protect the root and crown
from rot .
Forth,the plants were domesticated in closed bottles under the
nture light in greenhouse for 7d at first,then domesticated in
unclosed bottles for 3～7d.
Fifth,the plants were cultured the culture house, the moisture
was kept and sunlight was shaded moderately at the beginning stage
of transplanting.
At last, the disease and insects were controled, the water and
fertilication managements were improved after transplating. When the
grape seedlings growed for a period of time under nature conditions
then planted in the vineyard
号、泰山 12 号)以及无核葡萄与中国野生葡萄杂种(红光无核×山葡萄)的胚珠在不
同时间离体培养,并结合生理生化指标测定发现:不同的无核葡萄及其杂种胚败育的时
间不同,胚败育时,SOD、POD、CAT 酶活性下降、膜脂过氧化(MDA)水平上升,可溶性
糖与可溶性蛋白质降低,可溶性淀粉变化不大,进行胚挽救时,应根据不同的杂交组合,
在最佳时间即胚发育程度最高,SOD、POD、CAT 三种保护酶活性最高,MDA 水平最低时
取胚珠培养,效果较好。
对胚挽救获得的试管苗经不同时间的炼苗,测定叶片气孔,叶绿素含量及保护酶
(SOD)活性,发现随闭瓶开瓶炼苗时间的延长,气孔开张度逐渐降低,关闭指数增加,
叶绿素含量增加,保护酶(SOD)活性增加。这充分说明了分步炼苗的必要性,经过分
步炼苗,苗叶生长健壮,保护组织形成,气孔恢复自我调节功能,根系吸收能力增强,
从而提高了试管苗对外界环境条件的适应能力,有利于移栽成活率的进一步提高。并提
出了无核葡萄及其杂种胚挽救试管苗最佳配套移栽技术:
①培育优质健壮试管生根苗;
②移栽基质和苗根部消毒;
③ 蛭石或珍珠岩(上层)与 田间土(下层)分层移栽,下面土层提供根系生长
2 3
5 5
发育的营养,而上面沙层提高根基部透气性,可防止根茎部霉烂;
④温室自然光闭口炼苗 7d,过渡到开口炼苗 3～7d;
⑤移栽初期注意保湿和适度遮阳,培养室条件下培养。
同时移栽后加强病虫防治和水肥管理,在自然条件下生长,最后定植到田间。
Different seedless grapes and their hybrids had different embryo
abortive period. During embryo abortion,the activities of SOD, POD,
CAT decreased,the level of MDA the soluble protein and soluble sugar
increased and the change of soluble starch was small.So during embryo
rescue process,we should select the optimum time to culture ovules
according to different cross combinations,that is the embryo
development level was best,the activities of SOD, POD, CAT is highest
and the level of MDA is lowest.All these results were obtained through
culturing ovules in vitro of seedless grapes (self-pollinated
Sultanina,Sultanina×Olmo),seededgrapes (Beichun, Taishan-1,
Taishan-12) and pollinations between seedless grapes and Chinese
wild grapes (Flame Seedless×Heilongjiang Seedling) and destmining
the physiological and biochemical index.
The open degress and the index of leaf stomates,the content
of chlorophyll and the activities of protective enzyme (SOD) were
determined.After tissue culture plants obtained from embryo rescue
being domesticated through different time.The results showed: the
prolonging domesticatng time by closed-unclosed bottle,the open
degress of stomate dropped,the index of stomate close,the contents
of chlorophyll and the activities of protective enzymes (SOD)
incressed. These indicated that it is necessary to domesticate plants
step by step.And by this kind of domestication,the plants growed
stronghy,the protective tissue developed well,self-adjusting
function of stomate recovered and the absorption capacity of roots
increased. So the adaptability of the embryo rescue plants to
envierment is improved,thus it is benefit for transplanting survival
rate.In this study,the optimam transplanting technology of seedless
grapes and their hybrids was obtained:
First,good quality tissue culture plants with roots were
necessary.
Second,the transplanting matrix and the roots of plants must be
steriled.
Third,the optimam matrix is vermiculite or pearlit (upper
2
5
layer) and 3 nutrient soil (under layer) the nourishing soil can
5
provide the nourishment for root growing. While the vermiculite or
pearlite can improve capacity.Thus it can protect the root and crown
from rot .
Forth,the plants were domesticated in closed bottles under the
nture light in greenhouse for 7d at first,then domesticated in
unclosed bottles for 3～7d.
Fifth,the plants were cultured the culture house, the moisture
was kept and sunlight was shaded moderately at the beginning stage
of transplanting.
At last, the disease and insects were controled, the water and
fertilication managements were improved after transplating. When the
grape seedlings growed for a period of time under nature conditions
then planted in the vineyard
引文