[分享]食用菌菌种的制作与培养技术3
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt" class="MsoNormal"><b style="mso-bidi-font-weight: normal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">四、食用菌接种与培养技术<span lang="EN-US"><?xml:namespace prefix = o ns = "urn:schemas-microsoft-com:office:office" /><o:p></o:p></span></span></b></p><p style="MARGIN: 0cm="0cm"0cm="0cm"0pt" class="MsoNormal"><b style="mso-bidi-font-weight: normal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt"> </span></b><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">（一）、食用菌的接种技术；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt"> 接种是食用菌制种工作中的一项最基本的操作，无论是食用菌的传代，分离、鉴定还是进行食用菌的形态、生理等方面的研究都离不开接种操作。接种的关键是严格无菌操作，根据不同的目的、不同的菌类及同一菌类的不同菌种容器，接种方法都有所区别，但在无菌条件进行严格的无菌操作这一点是必须共同遵守的。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"42pt; mso-list: l1="l1"level1="level1"lfo1; tab-stops: list="list"42.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">１.</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">无菌操作规程<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l4="l4"level1="level1"lfo2; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（１）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种前对接种箱（室）进行清洁消毒，准备好接种工具。将待接的母种培养基或原种培养基或栽培种培养基放入接种箱内或室内，用药物熏蒸消毒，有条件也可用紫外线灯、净化设备消毒灭菌；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l4="l4"level1="level1"lfo2; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（２）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">换好清洁的工作衣，取少许药棉，蘸上<span lang="EN-US">75%</span>的酒精擦拭双手，菌种容器表面，工作台面及接种工具；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l4="l4"level1="level1"lfo2; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（３）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">点然酒精灯开始接种操作。酒精灯火焰周围<span lang="EN-US">8cm</span>半径范围的空间为无菌区，接种操作必须在无菌区内进行。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"42pt; mso-list: l1="l1"level1="level1"lfo1; tab-stops: list="list"42.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">２.</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种方法<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l5="l5"level1="level1"lfo3; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（１）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">母种接种<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">按上述要求做好无菌操作准备后，进行接种。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">ａ．用<span lang="EN-US">75%</span>酒精棉球涂擦工作人员的手和菌种试管外壁进行消毒，放入接种箱或接种室内；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">ｂ．用酒精棉涂擦接种针，再在酒精灯上烧红；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">ｃ．左手排握着菌种和斜面试管，右手握持接种针，用中指与无名指、无名指与小指；分别把菌种和斜面培养基上的试管棉塞拔开；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">ｄ．用接种钩伸入菌种管割取菌种，不宜过大，米粒大小，迅速转接入斜面培养基中央；将试管口在酒精灯火焰上灼烧一下；塞好试管棉塞；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">ｅ．接上菌种的斜面试管贴上标签，注明菌名和接种日期等。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt" lang="EN-US"><o:p> </o:p></span></p>
<p style="TEXT-INDENT: 12pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"59.95pt; mso-char-indent-count: 1.0; mso-para-margin-left: 5.71gd" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">整个转管过程应在酒精灯火焰<span lang="EN-US">8-10c</span>㎡</span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 宋体">范围的无菌区进行。转接后的试管，放入适宜的温度下培养至长满斜面。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l5="l5"level1="level1"lfo3; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（２）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 宋体">原种接种<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">要求做好无菌操作准备后，进行接种。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt; mso-list: l0="l0"level1="level1"lfo4; tab-stops: list="list"78.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">a.<span style="FONT: 7pt="7pt"'Times New="New"Roman'"> </span></span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">将灭过菌的菌种瓶、母种和接种钩等一起放入接种箱消毒后开始接种。母种接原种，一般一支斜面试管可接原种<span lang="EN-US">4</span>～<span lang="EN-US">6</span>瓶。接种时严格按无菌操作规程。双手要用<span lang="EN-US">75%</span>酒精棉球表面消毒，拔开母种棉塞，试管口用酒精灯火焰封口。接种钩要经过火焰灭菌。然后将母种（菌丝）连同培养基用接种钩分成<span lang="EN-US">4</span>～<span lang="EN-US">6</span>段，每一段母种迅速转移到原种培养基上。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt; mso-list: l0="l0"level1="level1"lfo4; tab-stops: list="list"78.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">b.<span style="FONT: 7pt="7pt"'Times New="New"Roman'"> </span></span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接完种后，将待培养的菌种瓶移入培养箱或光线暗淡的培养室，平卧或立在瓶架上发菌，根据各种菌类的生长要求，调整适宜的温度。培养初期逐日观察菌丝定植生长和杂菌污染情况。如接种<span lang="EN-US">5</span>天仍未见菌丝萌发则应及时补接母种，如发现杂菌应及时清除。同时要经常调瓶的位置，以利于菌丝生长一致，当菌长满瓶后及时使用。若暂时不用刻放低温、干燥、避光的环境下短期保存，一般棉子皮原种<span lang="EN-US">25</span>天左右长满瓶，麦粒原种<span lang="EN-US">15</span>天左右。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt; mso-list: l5="l5"level1="level1"lfo3; tab-stops: list="list"60.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（３）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">栽培种接种<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"60pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">要求做好无菌操作准备后，进行接种。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt; mso-list: l3="l3"level1="level1"lfo5; tab-stops: list="list"78.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">a.<span style="FONT: 7pt="7pt"'Times New="New"Roman'"> </span></span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">装箱 ：<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: 18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"59.95pt; mso-char-indent-count: 1.5; mso-para-margin-left: 5.71gd" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">将灭过菌的栽培袋（瓶），接种用的器具，原种等放入接种箱中。将待接的栽培袋（瓶）置于接种箱一端，袋横卧码好，瓶立码。然后进行空间消毒。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt; mso-list: l3="l3"level1="level1"lfo5; tab-stops: list="list"78.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">b.<span style="FONT: 7pt="7pt"'Times New="New"Roman'"> </span></span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种：<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: 18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"59.95pt; mso-char-indent-count: 1.5; mso-para-margin-left: 5.71gd" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">原种瓶口应用<span lang="EN-US">75%</span>酒精棉球擦拭，进行瓶口消毒。然后用火焰灭过菌的接种钩清理原种表皮的均被，再将长满菌丝的培养料挖成粒（块）状。在酒精灯火焰旁，解开袋绳结，用接种钩将原种颗粒（块）移植到栽培种的培养料袋（瓶）内。一般每瓶原种可接<span lang="EN-US">35</span>～<span lang="EN-US">50</span>袋（瓶）<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt; mso-list: l3="l3"level1="level1"lfo5; tab-stops: list="list"78.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">c.<span style="FONT: 7pt="7pt"'Times New="New"Roman'"> </span></span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">培养：<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"78pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接完种后，将待培养的菌种袋（瓶）置培养室发菌，在培养过程中定期向地面喷洒化学消毒药品，防止空气污染。接种后<span lang="EN-US">3</span>～<span lang="EN-US">10</span>天经常观察菌丝生长情况，菌种<span lang="EN-US">30</span>天左右即可长满。菌丝长满一周后，即可栽培。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -18pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"42pt; mso-list: l1="l1"level1="level1"lfo1; tab-stops: list="list"42.0pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">３.</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种注意事项<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"66.05pt; mso-list: l2="l2"level1="level1"lfo6; tab-stops: list="list"66.05pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（１）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种前要准备一些无菌棉塞，一起放入无菌室（箱内），以便在所用棉塞受潮时更换；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"66.05pt; mso-list: l2="l2"level1="level1"lfo6; tab-stops: list="list"66.05pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（２）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种时切勿使试管口、瓶口向上，切勿离开酒精灯火焰的无菌区进行；人在室内尽量少走动，减少空气流动扬起的灰尘、减少污染；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"66.05pt; mso-list: l2="l2"level1="level1"lfo6; tab-stops: list="list"66.05pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（３）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">接种时留下的污物，如用过的酒精棉、菌种碎屑等要及时清除，以免污染；<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: -36pt; MARGIN: 0cm="0cm"0cm="0cm"0pt="0pt"66.05pt; mso-list: l2="l2"level1="level1"lfo6; tab-stops: list="list"66.05pt" class="MsoNormal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt; mso-bidi-font-family: 新宋体" lang="EN-US"><span style="mso-list: Ignore">（４）</span></span><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt">如一次接种不同菌种时，要注意做好标记，以免搞混。<span lang="EN-US"><o:p></o:p></span></span></p>
<p style="TEXT-INDENT: 12.05pt; MARGIN: 0cm="0cm"0cm="0cm"0pt; mso-char-indent-count: 1.0" class="MsoNormal"><b style="mso-bidi-font-weight: normal"><span style="FONT-FAMILY: 新宋体; FONT-SIZE: 12pt" lang="EN-US"><o:p> </o:p></span></b></p>
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