CN106259460A - 一种提高胡萝卜中总糖和产量的复合菌剂及其应用 - Google Patents
一种提高胡萝卜中总糖和产量的复合菌剂及其应用 Download PDFInfo
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Abstract
本发明公开了一种提高胡萝卜中总糖含量和胡萝卜产量的复合菌剂及其应用，该复合菌剂由植物乳杆菌(Lactobacillus plantarum)P‑8和干酪乳杆菌(Lactobacillus casei)Zhang组成，这两种菌对胡萝卜的生长代谢有明显的影响，尤其可以提高胡萝卜总糖含量和胡萝卜产量，从而为胡萝卜食品的制备提供了较为理想的原料，最终改善了胡萝卜食品的口感，降低了胡萝卜食品加工中外源糖分的添加，提高了产品的品质。
Description
技术领域
本发明属于微生物技术领域，涉及植物乳杆菌(Lactobacillus plantarum)P-8和干酪乳杆菌(Lactobacillus casei)Zhang及其代谢产物组成的混合菌剂，尤其是一种能显著提高胡萝卜中总糖含量和胡萝卜产量的菌剂。
背景技术
胡萝(Daucus carrot)，又称甘荀，是伞形科胡萝卜，属于二年生草本植物。以肉质根作蔬菜食用，栽培历史在2000年以上。胡萝卜中含有蛋白质、脂肪、糖类化合物(蔗糖、葡萄糖)，粗纤维，钙、磷、铁，硫胺素，核黄素，尼克酰，抗坏血酸，挥发油(a-蒎烯，茨烯，月桂烯，柠檬烯)等成份。胡萝卜中的α，β，ρ胡萝卜素和多种类胡萝卜素是VA的来源，细胞壁中含有极丰富的果胶酸脂。
胡萝卜不仅是一种质脆味美、营养丰富的家常蔬菜，也有很好的医疗作用：美国科学家研究证实，每天吃两根胡萝卜，可使血中胆固醇降低10％-20％；每天吃三根胡萝卜，有助于预防心脏疾病和肿瘤；日本科学家发现，胡萝卜中的β-胡萝卜素能有效预防花粉过敏症、过敏性皮炎等过敏性疾病，且β-胡萝卜素能调节细胞内的平衡，从而使身体不易出现过敏反应；德国的一项研究表明，胡萝卜含抗氧化剂胡萝卜素，皮肤中抗氧化剂浓度高的人，皱纹明显比皮肤中抗氧化剂浓度低的人少；中医认为，胡萝卜味甘、性平，无毒，入肺、脾经，具有健脾消食，润肠通便，杀虫，行气化滞，补肝明目，清热解毒等功效；能治疗食欲不振、腹胀、腹泻、咳喘痰多、视物不明、小儿营养不良、麻疹、夜盲症、便秘、高血压、肠胃不适、饱闷气胀等，素有“小人参”之称。
当前，胡萝卜的营养价值已被越来越多的人认可和接受，除了传统作为家常蔬菜烹饪食用之外，胡萝卜精深加工也成为一种趋势，如专利CN104286337A公开了一种胡萝卜果脯的制备方法；专利CN101972018A公开了一种医用保健用胡萝卜浓缩汁及其制备方法；专利CN104012612A公开了一种富含胡萝卜的学生营养饼干。综上，胡萝卜越来越多的以多种形式被人们食用或利用，提高胡萝卜产量有助于提高胡萝卜的生产效益，而胡萝卜中糖分的提高有助于改善胡萝卜口感，同时降低胡萝卜食品加工中外源糖分的添加，提高产品的品质。
发明内容
本发明的目的在于公开一种植物乳杆菌(Lactobacillus plantarum)P-8和干酪乳杆菌(Lactobacillus casei)Zhang组成的混合菌剂在提高胡萝卜总糖含量和胡萝卜产量中的应用。
为了实现本发明的目的，发明人通过大量试验研究并不懈努力，最终获得了如下技术方案：
一种复合菌剂，该菌剂由植物乳杆菌(Lactobacillus plantarum)P-8和干酪乳杆菌(Lactobacillus casei)Zhang组成。
需要说明的是，本发明所采用的植物乳杆菌(Lactobacillus plantarum)P-8，分离自内蒙古传统自然发酵酸牛奶中，具有优异的抗胃肠道消化液耐受能力，能够存活于动物肠道内；该菌株已保藏于中国微生物菌种保藏管理委员会普通微生物中心，地址为中国北京市朝阳区北辰西路1号院3号，保藏时间为2012年06月28日，保藏号为CGMCC No.6312。另外，本发明所采用的干酪乳杆菌(Lactobacillus casei)Zhang，是从酸马奶中分离的耐酸和耐胆汁酸的益生菌菌株；该菌株于2006年4月21日保藏于中国微生物菌种保藏管理委员会普通微生物菌种保藏中心，保藏号CGMCC No.1697。
优选地，如上所述的复合菌剂，该菌剂由植物乳杆菌(Lactobacillus plantarumP-8)和干酪乳杆菌(Lactobacillus casei)Zhang按1：(0.5～3)的活菌数量比组成。
本发明人在筛选抑制胡萝卜病原菌的拮抗菌时意外发现，含有植物乳杆菌(Lactobacillus plantarum)P-8和干酪乳杆菌(Lactobacillus casei)Zhang的复合菌液对病原菌没有明显抑制作用，但对胡萝卜的生长代谢有明显的影响，尤其可以显著提高胡萝卜总糖含量和胡萝卜产量。基于此发现，本发明人通过接种于MRS培养液中培养发酵，将这两种菌制备成了复合菌剂。因此，本发明提供了上述复合菌剂的应用，即：上述的复合菌剂在提高胡萝卜中总糖含量中的应用；上述的复合菌剂在提高胡萝卜产量中的应用。以及，上述的复合菌剂在制备提高胡萝卜产量的生长液中的应用。
本发明涉及的复合菌剂可以采用如下方法制备：
(1)将所述干酪乳杆菌菌株(Lactobacillus casei)Zhang和植物乳杆菌菌株(Lactobacillus plantarum)P-8接种于MRS培养液，在37℃条件下培养12-24小时，经离心洗涤后，在菌体沉淀上加入8％-12％灭菌脱脂牛乳液，并调整其菌数为2.0×1010cfu/mL，混匀后倾注培养；
(2)将步骤(1)得到的菌株培养物接种于MRS培养液，在37℃条件下培养12-24小时，再接种8％-12％的灭菌脱脂牛乳液，在37℃条件下培养6-10小时，调整其菌数为3×109cfu/mL，pH值为4.4～4.6。
与现有技术相比，本发明涉及的复合菌剂由植物乳杆菌(Lactobacillusplantarum)P-8和干酪乳杆菌(Lactobacillus casei)Zhang组成，这两种菌对胡萝卜的生长代谢有明显的影响，尤其可以提高胡萝卜总糖含量和胡萝卜产量，从而为胡萝卜食品的制备提供了较为理想的原料，最终改善了胡萝卜食品的口感，降低了胡萝卜食品加工中外源糖分的添加，提高了产品的品质。
附图说明
图1为标准曲线，横坐标为蔗糖浓度(μg/ml)，纵坐标为吸光值。
具体实施方式
以下通过实施例形式对本发明的上述内容再作进一步的详细说明，但不应将此理解为本发明上述主题的范围仅限于以下的实施例，凡基于本发明上述内容所实现的技术均属于本发明的范围。另外，下述实施例中的实验方法，如无特别说明，均为常规方法。
实施例1：植物乳杆菌(Lactobacillus plantarum P-8)和干酪乳杆菌(Lactobacillus casei)Zhang在提高胡萝卜中总糖含量和胡萝卜产量的试验
实验地点：莱西店埠后屯(植物乳杆菌组、干酪乳杆菌组、复合菌组和空白对照组各1.7亩)
种植管理：棚规格长115m，宽10m，每棚起垄13垄，每垄上种植萝卜2行，株距5-6cm，每棚施基肥：硫酸钾复合肥(N:P:K为15:15:15)300斤，有机肥600-700斤，后期不追肥。
实验时间：2016.04.26-2016.6.23
实验作物：胡萝卜(品种为：孟德尔808)
使用量及方法：胡萝卜播种结束之后，立即随水滴灌微生物菌剂(实验组采用的菌剂中活菌总数含量大于109cfu/mL，复合菌组中两种菌的数量比为1:1)2升/棚，后期不在滴灌含菌水，而根据作物长势灌溉不含菌水，保证各处理灌溉量相同。
测定方法：
1)标准曲线的制作
取6支试管按表加入试剂，再按顺序加入1.0ml 90g/L苯酚溶液，摇匀，然后再加入5.0ml浓硫酸，摇匀，室温下静止30min。然后以空白为参比，在波长485nm处测吸光值。
2)总糖提取
从不同胡萝卜的不同部位称取胡萝卜组织1.0g置于研钵中，研磨成浆，加入少量蒸馏水转入到刻度试管中，重复洗涤3-5遍，用塑料膜封口于沸水中煮沸提取30min，待冷却后，将滤液过滤到100ml的容量瓶中，再将残渣回收到试管中再次煮沸10min，冷却过滤然后定容。
3)测定
取0.5ml样品加入1.5ml蒸馏水，然后步骤与制作标曲相同。测定时如果读数过高，可将样品稀释后再进行测定。注意切记对反应后的混合液进行稀释
计算公式：
M0---从标曲查得的蔗糖质量，μg；
V--样品提取液总体积，ml；100
N--样品提取液稀释倍数；10倍
Vs--测定时所取样品提取液体积，ml；0.5
M--样品质量.1.0
表1各处理胡萝卜总糖含量及总产量
与空白对照组比较，▲P＜0.01；与植物乳杆菌组比较，P＜0.01；与干酪乳杆菌组比较，■P＜0.01。
实施例2：植物乳杆菌(Lactobacillus plantarum P-8)和干酪乳杆菌(Lactobacillus casei)Zhang在提高胡萝卜中总糖含量和胡萝卜产量的试验
实验地点：莱西店埠后屯
实验时间：2016.04.26-2016.6.23
实验作物：胡萝卜(品种为：诺利红)
使用量及方法：胡萝卜播种之后，随水滴灌微生物菌剂(两种菌的数量比为1:1，活菌总数含量大于109cfu/mL)2升/棚，后期不再滴灌含菌水，而根据作物长势灌溉不含菌水，保证各处理灌溉量相同。
种植管理方式、总糖含量测定方法(高效液相法)：同实施例1
表2各处理胡萝卜总糖含量及总产量
与空白对照组比较，▲P＜0.05；与植物乳杆菌组比较，P＜0.05；与干酪乳杆菌组比较，■P＜0.05。
由表1中检测结果可知，在胡萝卜品种“孟德尔808”上，与空白对照组相比，复合菌组可显著提高胡萝卜中总糖的含量及胡萝卜产量(p<0.05)；相对于空白对照，复合菌组总糖含量提高了16.70％，胡萝卜产量提高了10.40％；表2中检测结果可知，在胡萝卜品种“诺利红”上，与空白对照组相比，复合菌组也显著提高了胡萝卜中总糖的含量及胡萝卜产量(p<0.05)；相对于空白对照，实验组总糖含量提高了15.24％，胡萝卜产量提高了13.79％。值得注意的是，无论是在胡萝卜品种“诺利红”上还是在胡萝卜品种“孟德尔808”上，复合菌组对总糖含量的提高程度相比各单菌组均有显著性差异(p<0.05)。
Claims (3)
1.一种复合菌剂在提高胡萝卜中总糖含量中的应用，所述的复合菌剂由植物乳杆菌(Lactobacillus plantarum P-8)和干酪乳杆菌(Lactobacillus casei)Zhang按1∶(0.5～3)的活菌数量比组成。
2.一种复合菌剂在提高胡萝卜产量中的应用，所述的复合菌剂由植物乳杆菌(Lactobacillus plantarum P-8)和干酪乳杆菌(Lactobacillus casei)Zhang按1∶(0.5～3)的活菌数量比组成。
3.一种复合菌剂在制备提高胡萝卜产量的生长液中的应用，所述的复合菌剂由植物乳杆菌(Lactobacillus plantarum P-8)和干酪乳杆菌(Lactobacillus casei)Zhang按1∶(0.5～3)的活菌数量比组成。
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