植物学报
植物學報
식물학보
ACTA BOTANICA SINICA
2001年
5期
461-468
,共8页
李一勤%小竹敬久%樱井直树%赵南明%刘强
李一勤%小竹敬久%櫻井直樹%趙南明%劉彊
리일근%소죽경구%앵정직수%조남명%류강
细胞壁%β-葡聚糖酶%麝香百合%花粉管%顶端生长%野尻酶素
細胞壁%β-葡聚糖酶%麝香百閤%花粉管%頂耑生長%野尻酶素
세포벽%β-포취당매%사향백합%화분관%정단생장%야고매소
从离体培养的麝香百合(LiliumlongiflorumThunb.)花粉管细胞壁中分离纯化出β-葡聚糖酶。抑制剂野尻酶素(nojirimycin)使该酶活性明显降低。若在百合花粉培养的初始或培养1h时加入3mmol/L野尻酶素,与对照相比较,花粉的萌发率分别被抑制99.6%和91.4%。若把3mmol/L野尻酶素分别在培养的不同时刻(0、1、1.5和2h)加入到液体培养基中,花粉管的生长都即刻受阻。此后,花粉管生长方向稍有改变,且花粉管直径略增,呈弯曲的非正常形态。用视频显微术跟踪记录在正常或含有不同浓度野尻酶素(0.003、0.03、0.3和3mmol/L)的半固体培养基中培养的花粉管的生长变化模式,发现该抑制剂明显降低花粉管的生长速率并影响其生长方向,而且花粉管直径、胞质内细胞器及囊泡的分布和顶端生长区的区域范围也呈现异常。野尻酶素对花粉管生长的抑制是可逆转的。这些结果表明,β-葡聚糖酶参与百合花粉的萌发和花粉管生长的调节。结合我们已报道的该酶的性质及作用方式,进而推测该酶通过降解1,3-/1,4-或1,3:1,4-β-葡聚糖,影响细胞壁的延展性,从而对花粉管顶端生长显示调节作用。
從離體培養的麝香百閤(LiliumlongiflorumThunb.)花粉管細胞壁中分離純化齣β-葡聚糖酶。抑製劑野尻酶素(nojirimycin)使該酶活性明顯降低。若在百閤花粉培養的初始或培養1h時加入3mmol/L野尻酶素,與對照相比較,花粉的萌髮率分彆被抑製99.6%和91.4%。若把3mmol/L野尻酶素分彆在培養的不同時刻(0、1、1.5和2h)加入到液體培養基中,花粉管的生長都即刻受阻。此後,花粉管生長方嚮稍有改變,且花粉管直徑略增,呈彎麯的非正常形態。用視頻顯微術跟蹤記錄在正常或含有不同濃度野尻酶素(0.003、0.03、0.3和3mmol/L)的半固體培養基中培養的花粉管的生長變化模式,髮現該抑製劑明顯降低花粉管的生長速率併影響其生長方嚮,而且花粉管直徑、胞質內細胞器及囊泡的分佈和頂耑生長區的區域範圍也呈現異常。野尻酶素對花粉管生長的抑製是可逆轉的。這些結果錶明,β-葡聚糖酶參與百閤花粉的萌髮和花粉管生長的調節。結閤我們已報道的該酶的性質及作用方式,進而推測該酶通過降解1,3-/1,4-或1,3:1,4-β-葡聚糖,影響細胞壁的延展性,從而對花粉管頂耑生長顯示調節作用。
종리체배양적사향백합(LiliumlongiflorumThunb.)화분관세포벽중분리순화출β-포취당매。억제제야고매소(nojirimycin)사해매활성명현강저。약재백합화분배양적초시혹배양1h시가입3mmol/L야고매소,여대조상비교,화분적맹발솔분별피억제99.6%화91.4%。약파3mmol/L야고매소분별재배양적불동시각(0、1、1.5화2h)가입도액체배양기중,화분관적생장도즉각수조。차후,화분관생장방향초유개변,차화분관직경략증,정만곡적비정상형태。용시빈현미술근종기록재정상혹함유불동농도야고매소(0.003、0.03、0.3화3mmol/L)적반고체배양기중배양적화분관적생장변화모식,발현해억제제명현강저화분관적생장속솔병영향기생장방향,이차화분관직경、포질내세포기급낭포적분포화정단생장구적구역범위야정현이상。야고매소대화분관생장적억제시가역전적。저사결과표명,β-포취당매삼여백합화분적맹발화화분관생장적조절。결합아문이보도적해매적성질급작용방식,진이추측해매통과강해1,3-/1,4-혹1,3:1,4-β-포취당,영향세포벽적연전성,종이대화분관정단생장현시조절작용。
β-Glucanases were found in the cell wall of Lilium longiflorumThunb. pollen tubes grown in vitro. The activity of β-glucanases was, in a certain extent, decreased by nojirimycin, an inhibitor of glucosidase. Pollen germination percentage reduced dramatically when nojirimycin was applied in the culture medium . In case that nojirimycin was added at 0 or 1 h after the onset of incubation, the inhibition rate was 99.6% and 91.4%, respectively. When 3 mmol/L of nojirimycin was applied in the liquid medium at 0, 1, 1.5 and 2 h after the onset of incubation, the growth of pollen tubes was interrupted, which resulted in the morphological change of the pollen tubes such as the newly grown portion of pollen tubes being bent, curved and swollen. Tracing the growth pattern of the individual pollen tube grown in semi-solid medium by video microscopy, the authors demonstrated that pollen tube growth rate was strongly inhibited by nojirimycin at concentrations ranged from 0.003 to 3 mmol/L. Moreover, the cytoplasmic arrangem ent and the morphology of the pollen tubes were also affected by nojirimycin. The growth inhi bition brought about by nojirimycin was reversible. These results indicated that β-glucanases, which degrade 1,3-β-glucan and/or 1,4-β-glucan or 1,3:1,4-β-g lucan constructed in the cell wall, are involved in pollen germination and pollen tube growth. It provides new insight into an understanding of the contribution of β-glucanases to the cell wall extensibility and the crucial role of cell wall in regards to the regulation of pollen tube growth.