阿拉伯半乳糖蛋白对喷鼻蕉悬浮细胞胚性状态的影响_小论文论文-by_gnksguybb[优质文档]

阿拉伯半乳糖蛋白对喷鼻蕉悬浮细胞胚性状态的影响_小论文论文-by_gnksguybb[优质文档] 阿拉伯半乳糖蛋白对香蕉悬浮细胞胚性状态的影响_小论文论文-By_gnksguybb 闃挎媺浼 崐涔崇硸铔嬬櫧瀵归 钑夋偓娴 粏鑳炶儦鎬х姸鎬佺殑褰卞搷_灏忚 鏂? 璁烘枃瀵艰 :锛氶噰鐢ㄧ伀绠 惣鑴傜硸鍑濊兌鐢垫吵娉曟祴瀹氫簡鍩瑰吇15 d鍚庣殑棣欒晧鎮 诞缁嗚優娑蹭綋鍩瑰吇鍩轰腑鐨勯樋鎷変集鍗婁钩绯栬泲鐧斤紙arabinogalactan-proteins, AGPs锛夌殑鍚 噺锛屽彂鐜板叿鏈変綋鑳氬彂鐢熻兘鍔涚殑璐?晧銆佸ぇ钑夊拰榫欑墮钑?lsquo;杩囧北棣?rsquo;鍝佺 鐨勮儦鎬ф偓娴 粏鑳烇紙embryogenicsuspension cells, ECS锛夊煿鍏诲熀涓 殑AGPs鍚 噺鍒嗗埆杈惧埌36銆?8銆?6 µg/mL锛岃?屽湪涓嶈兘杩涜 浣撹儦璇卞 鐨勯 鑺借晧‘濞佸粔鏂?rsquo;鍝佺 鐨勯潪鑳氭?ф偓娴 粏鑳烇紙non-embryogenicsuspension cells, NECS锛夌殑鍩瑰吇鍩轰腑妫?娴嬩笉鍒癆GPs锛岃 鏄庢偓娴 粏鑳炲垎娉屽埌鍩瑰吇鍩轰腑鐨凙GPs鍚 噺涓庨 钑夋偓娴 粏鑳炵殑鑳氭?х姸鎬佹湁涓?瀹氱殑鍏崇郴銆傚湪棣欒晧ECS鐨勬偓娴 煿鍏昏繃绋嬩腑锛屾坊鍔犱笉鍚屾祿搴,殑ßGlcY璇曞墏 锛?szlig;-glucosyl Yariv 璇曞墏, 涓?绉嶈兘涓嶢GPs鐗瑰紓鎬х粨鍚堝苟寮曡捣AGPs娌夋穩鐨勪汉宸ュ悎鎴愬寲瀛,墿璐 級 鍒版偓娴 煿鍏诲熀涓 彲寮曡捣鑳氭?х粏鑳炰笌闈炶儦鎬х粏鑳?锛圗CS/NECS锛?鐨勬瘮鍊间笅闄嶏紱鍦ㄩ 钑夌殑ECS浣撹儦璇卞 杩囩?涓 紝娣诲姞涓嶅悓娴撳害ßGlcY璇曞墏鍒颁綋鑳氳 瀵煎煿鍏诲熀涓 紝寮曡捣浣撹儦鍙戠敓棰戠巼涓嬮檷銆傝繖浜涚粨鏋滆〃鏄庯紝AGPs鍦ㄩ 钑夋偓娴 粏鑳炵殑鑳氭?т繚鎸佽繃绋嬩腑璧峰埌閲嶈 鐨勪綔鐢 紝骞朵笖鍏锋湁鍓傞噺鏁堝簲銆? 璁烘枃鍏抽敭璇嶏細闃挎媺浼 崐涔崇硸铔嬬櫧(AGPs)锛岄 钑夛紝鎮 诞缁嗚優 銆?銆?棣欒晧绂讳綋缁嗚優鑳氭?х殑鎭? 鍜屼繚鎸佹槸鍐冲畾鍏 跺啀鐢熻兘鍔涚殑鍏抽敭锛屼篃鏄 钑夌敓鐗?妧鏈 偛绉嶇殑閲嶈 鍓嶆彁銆備絾鏄 紝鑳氭?у弽搴斾綆銆佸懆鏈熼暱銆佽儦鎬ф槗涓?け绛夐棶棰樹竴鐩村埗绾,潃棣欒晧楂樻晥鑳氭?ф偓娴 粏鑳炲啀鐢熶綋绯荤殑寤虹珛鍙婂叾鍦ㄧ敓鐗?妧鏈 偛绉嶄腑鐨勫簲鐢ㄣ?傚洜姝わ紝濡備綍鏈夋晥鍦颁績浣块 钑夌 浣撶粏鑳炶儦鎬х殑鎭? 骞舵湁鏁堝湴淇濇寔锛屾垚涓洪 钑夌敓鐗?妧鏈 彂灞曠殑鏍稿績闂 銆? 銆?銆?AGPs锛圓rabinogalactan-proteins锛岄樋鎷変集鍗婁钩绯栬泲鐧斤級鍏锋湁缁嗚優闂翠俊鍙风殑浣滅敤銆傜爺绌惰〃鏄庡皬璁烘枃锛屼粠涓嶅悓妞嶇墿涓 垎绂昏幏寰楃殑AGPs瀵逛簬缁嗚優鐨勮儦鎬ф仮澶嶅叿鏈夌浉浼肩殑鐢熺悊鍔熻兘銆侺etarte 绛夊彂鐜伴噰鐢ㄩ樋鎷変集鑳跺彲浠ヤ唬鏇跨湅鎶ゅ煿鍏伙紙鍏卞煿鍏伙級淇冭繘灏忛害锛圱riticum aestivum L.锛夊皬瀛?瓙姣嶇粏鑳炲彂鑲叉垚瀹屾暣鐨勬 鐗 1]锛汦gertsdoter 鍜?Von Aronld浠庢尓濞佷簯鏉夛紙Picea abies锛夌 瀛愪腑鎻愬彇鐨凙GPs鍙 互淇冭繘鎸 ?浜戞潐鎮 诞缁嗚優鐨勪綋鑳氬彂鑲瞇2]銆備粠鑳?悵鍗滐紙Daucuscarota锛夌 瀛愪腑鍒嗙 鐨凙GPs鍙 互淇冭繘鑳?悵鍗滄偓娴 粏鑳炵殑浣撹儦鍙戠敓[3,4]銆侹reuger 绛夌爺绌跺彂鐜颁粠鑳?悵鍗滅 瀛愪腑鑾峰緱鐨凙GPs鍙 互淇冭繘浠欏 鏉ワ紙Cyclamen persicum锛夊湪鍩瑰吇杩囩?涓 儦鎬х粏鑳炲洟鐨勫 鍔狅紝璁や负AGPs鐨勬椿鎬ф病鏈夊搧绉嶇壒寮傛? 5]銆傝繖浜涘疄楠岄兘琛ㄦ槑锛屼粠涓嶅悓鐨勬 鐗?潗鏂欒幏寰楃殑AGPs鍏锋湁淇冭繘鍏跺畠妞嶇墿浣撹儦鍙戠敓鐨勮兘鍔涳紝AGPs鍦ㄥ緢澶氭 鐗?粏鑳炶儦鎬х殑淇濇寔涓婂叿鏈夐噸瑕佺殑浣滅敤銆傛潕浣崇瓑鐨勭爺绌跺彂鐜板ぇ钑夛紙Musa paradisiaca Linn. cv. Da JiaoABB锛夌殑闈炶儦鎬ф偓娴 粏鑳炰笌璐?晧[Musa acuminata cv. Mas 锛圓A锛塢鐨勮儦鎬ф偓娴 粏鑳炴偓娴 煿鍏绘恫鐨凙GPs鍚 噺宸 埆寰堝ぇ[6]銆? 銆?銆??GlcY-璇曞墏鏄 竴绉嶄笌AGPs鐗瑰紓鎬х粨鍚堢殑鍖栧 璇曞墏锛屽叿鏈夌敓鐗?椿鎬э紝鍦ㄨ繘琛孉GPs鍔熻兘鐨勭爺绌 舵椂閫氬父閲囩敤娣诲姞?GlcY-璇曞墏鐨勬柟娉曡繘琛屻?傚湪鑿婅妺锛圕ichorium锛夎儦鎬ф偓娴 粏鑳炵殑鐮旂?涓 紝鍙戠幇娣诲姞?GlcY-璇曞墏浼氬 鑷磋強鑺嬫偓娴 粏鑳炵殑鑳氭?т抚澶盵7]銆傛湰鏂囧 鍏锋湁鑳氭?у拰涓嶅叿澶囪儦鎬х殑棣欒晧鎮 诞缁嗚優鐨勬恫浣撳煿鍏诲熀涓 殑AGPs鍚 噺杩涜 浜嗘祴瀹氾紝骞堕?氳繃瀵硅儦鎬ф偓娴 粏鑳炵殑鍩瑰吇鍩轰腑娣诲姞?GlcY-璇曞墏鐨勬柟娉曪紝鐮旂?AGPs鍦ㄩ 钑夎儦鎬х殑淇濇寔涓婃墍璧风殑浣滅敤銆? 銆?銆?1鏉愭枡涓庢柟娉? 銆?銆?1.1 鏉愭枡 銆?銆?璐?晧[Musa acuminata cv. Mas 锛圓A锛塢锛岄緳鐗欒晧‘杩囧北棣?rsquo;鍝佺 锛圡usa AAB Silk cv. ‘Guoshanxiang’锛夛紝澶ц晧[Musa paradisiaca Linn. cv. Da Jiao锛圓BB锛塢锛岄 鑺借晧‘濞佸粔鏂?rsquo;鍝佺 锛圡usa AAA cv. Williams锛夌殑鏈 垚鐔熼泟鑺卞簭灏忚 鏂囷紝鍙栬嚜骞夸笢鐪佸啘涓氱 瀛，櫌鏋滄爲鐮旂?鎵?寤虹珛鐨?ldquo;鍥藉 鏋滄爲绉嶈川-骞垮窞鑽旀灊棣欒晧鍦?rdquo;銆? 銆?銆?鍙傜収榄忓渤鑽,瓑 [8]銆佹潕鍝茬瓑 [9] 鐨勬柟娉曪紝浠ヨ础钑夈?佸ぇ钑夈?侀 鑺借晧‘濞佸粔鏂?rsquo;鐨勬湭鎴愮啛闆勮姳涓哄 妞嶄綋寤虹珛鍧囪川鐨勭粏鑳炴偓娴 郴銆傞緳鐗欒晧‘杩囧北棣?rsquo;鐨勮儦鎬х粏鑳炴偓娴 郴锛圗CS锛夌殑寤虹珛鍙傜収榄忓渤鑽,瓑鐨勬柟娉曡繘琛孾10]銆傜粏鑳炴偓娴 郴淇濇寔鍦?2娑蹭綋鍩瑰吇鍩篬11]涓 ?傛偓娴 煿鍏讳繚鎸佸湪鎸佺画110 rpm 鐨勬棆杞 憞搴婏紝28±1 鈩冦?侀粦鏆楁潯浠朵笅杩涜 銆? 銆?銆?1.2棣欒晧鎮 诞缁嗚優浣撹儦鍙戠敓鑳藉姏鐨勬 楠? 銆?銆?灏嗘偓娴 粏鑳炵洿鎺ユ帴绉嶅埌M3鍩瑰吇鍩篬11]锛岃 绠椾綋鑳氬彂鐢熼 鐜囦綔涓鸿 閲忔偓娴 粏鑳炶儦鎬х殑鏍囧噯銆傝 瀵间綋缁嗚優鑳氱殑鍙戠敓M3鍩瑰吇鍩虹粍鎴愪负锛歋H 鍩瑰吇鍩篬12]涓 殑澶ч噺鍏冪礌鍜屽井閲忓厓绱犲強鍏堕搧鐩? MS缁寸敓绱燵13]+ 4.5 μmol/L 鐢熺墿绱?+ 680μmol/L 璋锋皑閰拌兒+ 2 μmol/L鑴 皑閰?+ 100mg/L 楹，娊姘磋В鐗?+ 1.1 μmol/L NAA + 0.2 μmol/L 鐜夌背绱?+ 0.5 μmol/L 婵?鍔ㄧ礌 + 0.7μmol/L 2-浜岀敳鍩轰笝鐑 槍鍛? 29 mmol/L 涔崇硸 + 130mmol/L 钄楃硸 + 2 g/L Gelrite銆傚叿浣撴柟娉曞弬鐓ч瓘宀宠崳鐨勬柟娉曡繘琛孾8],浣撹儦璇卞 棰戠巼=浣撹儦鎬绘暟/ml PCV ECS銆? 銆?銆?1.3 棣欒晧鎮 诞缁嗚優鍩瑰吇娑蹭腑鐨凙GPs鐨勫惈閲忔祴瀹? 銆?銆??GlcY-璇曞墏鐨勫埗澶囧弬鐓 ariv绛夌殑鏂规硶[14]杩涜 锛涙偓娴 粏鑳炴恫浣撳煿鍏诲熀涓 殑AGPs娴撳害閫氳繃鐏 鐞艰剛绯栧厤鐤 數娉虫柟娉曡繘琛屾祴瀹歔15]銆? 銆?銆?1.4 ?GlcY-璇曞墏瀵归 钑夋偓娴 粏鑳炵敓闀跨姸鎬佸拰浣撹儦鍙戠敓鑳藉姏鐨勫奖鍝? 銆?銆?灏?GlcY-璇曞墏鎸?銆?銆?0銆?0銆?0 銆?0 μM鐨勬祿搴，姞鍏ュ埌M2娑蹭綋鎮 诞鍩瑰吇鍩轰腑锛岄緳鐗欒晧‘杩囧北棣?rsquo;鑳氭?ф偓娴 粏鑳炴寜鐓?.5%娴撳害鍒嗗埆鎺ョ 鍒颁笂杩板惈鏈変笉鍚屾祿搴?GlcY-璇曞墏鐨勫煿鍏诲熀涓 ??5 d鍚庤 绠楁偓娴 粏鑳炵殑PCV锛坧acked cell volume锛岀粏鑳炲瘑瀹炰綋绉 級锛屽苟鍦ㄦ樉寰 暅瑙嗛噹涓嬬粺璁?儦鎬у拰闈炶儦鎬х粏鑳烇紙ECS/NECS锛夌殑姣斾緥銆? 銆?銆?灏?GlcY-璇曞墏鎸夌収涓嬪垪娴撳害鍔犲叆鍒癕3鍩瑰吇鍩轰腑: 0銆?銆?0銆?0銆?0 μM銆?0 μM锛屽皢鎺ョ 缁т唬7-10 d鐨勫ぇ棰楃矑榫欑墮钑?lsquo;杩囧北棣?rsquo;鍝佺 鍝佺 鎮 诞缁嗚優鐢ㄤ簬杩涜 浣撹儦璇卞 锛?0 d鍚庣粺璁′綋鑳氬彂鐢熺巼鍜岀粏鑳炴?讳綋绉 紝浣撹儦璇卞 棰戠巼=浣撹儦鎬绘暟/ml PCV ECS銆? 銆?銆?2缁撴灉涓庡垎鏋? 銆?銆?2.1涓嶅悓鍩哄洜鍨嬮 钑夋偓娴 粏鑳炲煿鍏绘恫涓 殑 AGPs鐨勫惈閲忎笌浣撹儦鍙戠敓鐨勫叧绯? 銆?銆?琛?涓嶅悓鍩哄洜鍨嬮 钑夋偓娴 粏鑳炵殑浣撹儦鍙戠敓鑳 藉姏涓庡煿鍏诲熀涓瑼GPs鍚 噺娴嬪畾 銆?銆?Table 1 Quantification of thefrequency of somatic embryogenesis and AGPs content in liquid medium of cellsuspensions of different genotype of banana 銆?銆? 涓嶅悓鍩哄洜绫诲瀷棣欒晧 Different genotype of banana AGPs鍚 噺锛?mu;g/mL锛? AGPs content锛?mu;g/mL锛? 浣撹儦鍙戠敓棰戠巼锛?05浣撹儦/mL PCV ECS) Frequency of somatic embryogenesis 锛?105 embryos/mL PCV ECS) 榫欑墮钑?lsquo;杩囧北棣?rsquo;Musa AAB Silk cv. ‘Guoshanxiang’ 56±2.1 3.7±0.7 澶ц晧Musa paradisiaca Linn.cv.Da Jiao锛圓BB) 48±1.9 2.6±0.5 璐?晧Musa acuminata cv. Mas 锛圓A) 36±1.5 2.8±0.2 棣欒娊钑?lsquo;濞佸粔鏂?rsquo;Musa AAA cv. Williams 0 0 琛ㄤ腑鏁版嵁鏄 笁涓 噸澶嶅疄楠岀殑骞冲潎鍊?± 鏍囧噯璇疍ata represent average ± S.E. of three replicates. 銆?銆?浠庡 鍚勭 棣欒晧鎮 诞缁嗚優杩涜 浣撹儦璇卞 鐨勭粨鏋滄樉绀猴紙琛?锛夛紝榫欑墮钑?lsquo;杩囧北棣?rsquo;銆佽础钑夈?佸ぇ钑夌殑浣撹儦鍙戠敓鑳藉姏杈冨己锛岀浉搴旂殑鎮 诞鍩瑰吇娑蹭腑鐨凙GPs鍚 噺杈冮珮锛涢 鑺借晧‘濞佸粔鏂?rsquo;鐨勬偓娴 粏鑳炲熀鏈 病鏈変綋鑳氬啀鐢熻兘鍔涘皬璁烘枃锛屾偓娴 粏鑳炲煿鍏绘恫涓 篃妫?娴嬩笉鍒扮殑AGPs鐨勫瓨鍦ㄣ?傚洜姝わ紝鎮 诞鍩瑰吇娑蹭腑鐨凙GPs瀛樺湪鍙 綔涓烘 娴嬮 钑夋偓娴 粏鑳炶儦鎬х姸鎬佺殑涓?涓 綔鍦ㄦ寚鏍囥?? 銆?銆?2.2 ?GlcY-璇曞墏瀵归緳鐗欒晧‘杩囧北棣?rsquo;鑳氭?ф偓娴 粏鑳炵敓闀跨姸鎬佸拰浣撹儦鍙戠敓鑳藉姏鐨勫奖鍝? 銆?銆? 銆?銆?鍥? ?GlcY璇曞墏瀵归緳鐗欒晧‘杩囧北棣?rsquo;鎮 诞缁嗚優鐘舵?佺殑褰卞搷 銆?銆?a瀵圭収鑳氭?ф偓娴 粏鑳烇紝bar = 50 μm銆俠鐢?0 μM ?GlcY璇曞墏澶勭悊鍚庣殑鑳氭?ф偓娴 粏鑳? 銆?銆?Fig. 1-14 Effects of ?GlcY reagenton cell suspension of cv. Guoshanxiang (AAB) 銆?銆?a Embryogenic cellsuspension, bar = 50 μm. b Embryogenic cell suspension treated with 40 μM?GlcY, bar = 50 μm. 銆?銆?琛? ?GlcY-璇曞墏瀵归緳鐗欒晧‘杩囧北棣?rsquo;鑳 氭?ф偓娴 粏鑳炵敓闀裤?佺粏鑳炵姸鎬佸拰浣撹儦鍙戠敓鐨勫奖 鍝? 銆?銆?Table 2 Effects of ?GlcY on growth of ECS, cell statusand somatic embryogenesis of cv. Guoshanxiang 锛圓AB) 銆?銆? ?GlcY 娴撳害 锛?mu;M) Concentration of ?GlcY 锛?mu;M) 鎮 诞缁嗚優澧炲姞閲?锛坢L PCV) Added value of cell suspensions 锛坢l PCV) 鑳氭?х粏鑳炰笌闈炶儦鎬х粏鑳炵殑姣斿 ??EMs/NEMs 浣撹儦鍙戠敓棰戠巼锛?03涓? ml PCV ECS锛塅requency of somatic embryogenesis 锛?03 somatic embryos/ml PCV ECS) 0 2.9±0.20 a 11.90±1.26 a 15.5±1.41 a 5 2.7±0.15 a 1.42±0.11 bc 11.9±1.52 b 10 2.8±0.13 a 0.67±0.07 bcd 6.0±0.48 c 20 2.5±0.19 a 0.40±0.02 bcd 5.0±0.23 c 40 2.9±0.14 a 0.19±0.02 bcd 3.0±0.19 cd 60 2.9±0.22 a 0.03±0.01 e 0.2±0.01 e 瀹為獙缁撴灉鏄剧ず锛屽湪榫欑墮钑?lsquo;杩囧北棣?rsquo;鐨勮 儦鎬ф偓娴 粏鑳炲煿鍏诲熀涓 坊鍔犲悇绉嶆祿搴,殑?GlcY-璇 曞墏鍚庯紝缁忚繃15 d鐨勫煿鍏伙紝缁嗚優澧炴畺閲忔棤鏄捐憲鐨 勫彉鍖栵紝涔熷氨鏄 锛屾坊鍔?GlcY-璇曞墏涓嶅奖鍝嶇粏鑳炵 殑鍒嗚 鍜岀敓闀匡紙琛?锛夈?備絾鏄 樉寰 暅瑙傚療鍙戠幇锛 屼笌瀵圭収鑳氭?ф偓娴 粏鑳炵浉姣旓紙鍥?-a锛夛紝缁忚 繃?GlcY-璇曞墏澶勭悊鍚庯紝鍏锋湁鑳氭?х壒寰佺殑缁嗚優鍙婄粏 鑳炲洟鏁伴噺鏄捐憲涓嬮檷锛堝浘1-b锛岃〃2锛夊皬璁烘枃锛岀粷 澶у 鏁扮殑缁嗚優杞 彉涓洪暱鏉″舰锛屽唴鍚 墿鍚 噺闄嶄 綆鐨勫吀鍨嬬殑闈炶儦鎬х粏鑳炵姸鎬侊紝杩欎簺缁嗚優涓嶅叿澶 囦綋鑳氬彂鐢熻兘鍔涳紝璇存槑娣诲姞?GlcY-璇曞墏浣跨粏鑳炵姸 鎬佷粠鑳氭?х姸鎬佽浆鍙樹负闈炶儦鎬х姸鎬併?? 銆?銆?鍦?3鍩瑰吇鍩轰腑娣诲姞鍚勭 娴撳害鐨?GlcY鍚庯紝鍙戠 幇榫欑墮钑?lsquo;杩囧北棣?rsquo;鍝佺 缁嗚優鍥?湪M3鍩瑰吇 鍩轰笂缁х画澧炴畺锛?0 d鍚庯紝鍚勭 澶勭悊鐨勭粏鑳炴?讳綋 绉 棤鏄庢樉鐨勫樊寮傘?備絾鏄 紝缁忚繃45d鐨勫煿鍏伙紝鎮 诞缁嗚優鐨勪綋鑳氬彂鐢熼 鐜囬殢鐫??GlcY-璇曞墏娴撳害鐨勫 楂樿?屾樉钁椾笅闄嶃?傚綋?GlcY-璇曞墏娴撳害杈惧埌60 μM鏃 讹紝浣撹儦鍙戠敓棰戠巼宸茬粡闈炲父浣庛?傜粨鏋滆〃鏄庯 紝?GlcY瀵?rsquo;杩囧北棣?rsquo;棣欒晧鑳氭?х粏鑳炵殑鍒嗚 鍜屽 闀挎棤鏄捐憲褰卞搷锛屼絾鎶戝埗缁嗚優浣撹儦鍙戠敓鑳藉 姏灏忚 鏂囷紝涓斿叿鏈夊墏閲忔晥搴斻?? 鍙傝?冩枃鐚? [1] 鍙跺瓙锛岃繘锛庡疄鐜癧J]锛庤 绠楁満宸ョ?涓庣 瀛,紝 2009锛?(07)锛?1-64. [2] 鍩轰簬MATLAB 瀹炵幇鐨勬寚绾瑰浘鍍忛 澶勭悊[J]锛?008 锛?4(04)锛?36-856. [3] [4] [5] By-gnksguybb [6] 鏉?瀹侊紟鎸囩汗澧炲己绠楁硶鐮旂?[D]锛庢祹鍗楋細灞变笢 澶у 锛?007. [7] 椹 壃褰 紟鍩轰簬Gabor 婊ゆ尝鐨勬寚绾瑰浘鍍忓 寮篬J]. 寰 绠楁満淇?伅锛?009锛?5(15)锛?05-306. [8] 鍚?閿愶紟闈?悜浣庤川閲忔寚绾圭殑鍥惧儚澧炲己绠楁硶鐮旂 ?[D]锛庨噸搴嗭細瑗垮崡鏀挎硶澶у 锛?008.