测定植物离体根比生长速率的装置和方法 In vitro determination of plant root growth rate than the apparatus and method

技术领域 FIELD

[0001] 本发明涉及生物工程，具体涉及一种测定植物离体根比生长速率的装置和方法。 [0001] The present invention relates to biotechnology, specifically to a root plant body measuring specific growth rate from the apparatus and method. 背景技术 BACKGROUND

[0002] 我国是世界上药用植物资源最丰富的国家之一，但随着市场需求量的增加而造成了对药用植物资源的超量采挖，野生植物资源蕴藏量和产量普遍存在着下降趋势。 [0002] China is the world's most abundant resources of medicinal plants of the country, but with the increase in market demand caused by the excess of medicinal plant resources excavation, wild plant resources reserves and production prevails downward trend. 目前药用植物资源主要来源于大田种植。 Currently medicinal plant resources mainly from field planting. 大田栽培一方面受“天时地利”的影响，产量质量波动大；另一方面不可避免地带来农药残留污染等问题。 Field cultivation on the one hand by the "right place" the impact of large fluctuations in output quality; on the other hand inevitably bring pesticide pollution and other issues. 植物组织细胞生物反应器培养具有生长迅速，易于控制的特点，从而可以逐渐取代大田栽培满足市场的需要。 Cell bioreactor plant tissue culture has grown rapidly, easy to control, so you can gradually replace field cultivation to meet the needs of the market.

[0003]由于大部分药用植物以根入药，所以离体根培养是药用植物资源再生的热点之一。 [0003] Since most medicinal plant root used as medicine, so vitro root cultures is one of the hot-renewable resources of medicinal plants. 离体根生物反应器培养时，生长速率是衡量培养条件和反应器性能的一个重要指标。 When in vitro root bioreactor, the growth rate is an important indicator of the culture conditions and reactor performance. 为了实时了解植物离体根的生长规律，首先要画出其生长曲线从而算出比生长速率，即每天生长的量。 For real-time understanding of plant roots from growing body of law, the first to draw the growth curve in order to calculate the specific growth rate, that is growing daily amount. 植物悬浮细胞培养时，细胞会均一地分散到液体培养基中，所以每天只要取出一小部分细胞溶液，按比例就可以算出整个反应器中细胞的密度，从而得到比生长速率。 When the plant suspension cell cultures, cells uniformly dispersed in a liquid medium, so long as the cell solution was taken out a small part of a day, can be calculated in proportion to the density of the cells throughout the reactor, thereby to obtain the specific growth rate.

[0004] 离体根在反应器中生长时会相互缠绕逐渐形成一个团，不能象离体细胞那样形成均一的溶液，所以取出的样品中不含离体根只有培养液，无法算比生长速率。 [0004] When the in vitro growth of roots in the reactor will gradually wound around each other to form a group, that can not be like cells in vitro to form a uniform solution, so that the sample taken from the body does not contain only the root culture medium, the specific growth rate can not be calculated . 目前反应离体根生长情况的指标主要是平均生长速率，即V =(终浓度-接种浓度）/生长时间，但有关比生长速率的计算还未见报道。 Currently reaction in vitro root growth of the main indicators is the average growth rate, ie V = (final concentration - inoculum) / growth time, but about the specific growth rate calculation has not been reported.

发明内容 SUMMARY

[0005] 本发明的目的，就是为了提供一种测定植物离体根比生长速率的装置和方法。 [0005] The present invention is to provide a specific growth rate determined stumps apparatus and method for plant away.

[0006] 为了达到上述目的，本发明采用了以下技术方案：一种测定植物离体根比生长速率的装置，包括顺序连接连通的离体根生物反应器、第一刻度管、虹吸管、废液瓶和第二刻度管；所述虹吸管的一端与第一刻度管的下部连接连通，虹吸管的另一端插入废液瓶内；所述第二刻度管与废液瓶的下部连接连通。 [0006] To achieve the above object, the present invention adopts the following technical scheme: A method for determining in vitro plant root specific growth rate, comprising sequentially connected communication excised roots bioreactor, a first calibration tube, siphon, waste liquid bottle and a second calibration tube; proximal end of said siphon tube is connected to the first scale communication, the other end of the siphon tube into the waste bottle; the lower portion of the second calibration tube connection communicating with the waste container.

[0007] 上述测定植物离体根比生长速率的装置，其中，所述第一刻度管上连接有带空气过滤器的通气管，所述废液瓶上连接有带空气过滤器的通气管。 [0007] The determination of in vitro plant root specific growth rate means, which is connected with the air filter on the first calibration tube snorkel, with a snorkel is connected to the air filter on the waste container .

[0008] 上述测定植物离体根比生长速率的装置，其中，所述第一刻度管的精度为0.1毫升；所述第二刻度管的精度为I毫升。 [0008] The determination of in vitro plant root specific growth rate means, wherein, the accuracy of the first calibration tube is 0.1 ml; accuracy of the second calibration tube is I ml.

[0009] 一种用上述测定植物离体根比生长速率的装置测定植物离体根比生长速率的方法，包括如下步骤： [0009] An in vitro by the measurement of plant root specific growth rate means the plant roots than in vitro growth rate, comprising the step of determining:

[0010] 步骤一，取多份不同体积的植物离体根，分别浸泡到装有培养液的量筒中，根据排开的水的体积，得出每份植物离体根的体积； [0010] Step a, to take multiple different volumes vitro plant roots were immersed in a graduated cylinder containing the culture medium in accordance with of the displaced volume of water, the volume of each plant obtained in vitro root;

[0011] 步骤二，在60°C下烘干植物离体根，称量得其干重； [0011] Step two, at 60 ° C under dry vitro plant roots, weighing lent dry weight;

[0012] 步骤三，以植物离体根的体积为横坐标，重量为纵坐标作图，并算出回归系数，即单位体积的植物离体根所对应的重量；[0013] 步骤四，把植物离体根样品接种到离体根生物反应器中培养，每天记录植物离体根排开水的体积； [0012] Step three, in order to plant roots from the volume of the body as the horizontal, the vertical axis is plotted weight, and calculate regression coefficients per unit volume of the plant roots in vitro corresponding weight; [0013] Step four, the plant Isolated Root Isolated Root samples inoculated into the bioreactor culture, recorded daily in vitro root plant row water volume;

[0014] 步骤五，将植物离体根排开水的体积与步骤三算出的回归系数相乘，得出相应植物离体根的干重； [0014] Step five, the plant discharge water from the body of the root volume calculated in step three regression coefficients are multiplied to give the corresponding vitro plant root dry weight;

[0015] 步骤六，作出生长曲线图； [0015] Step six, making growth curve;

[0016] 步骤七，结合生长曲线图，根据公式 [0016] Step seven, combined with the growth curve, according to the formula

计算出比生长速率， Calculated specific growth rate,

[0017] 式中，X为终浓度，t为培养时间，X和Xtl为接种浓度，U为比生长速率； [0017] wherein, X is the final concentration, t is incubation time, X and Xtl as inoculum concentration, U for the specific growth rate;

[0018] 所述终浓度为培养结束时离体根的干重克数与培养液体积毫升数之比； [0018] The final concentration of the number of grams of dry weight when cultured in vitro root end with a few ml of the culture liquid volume ratio;

[0019] 所述接种浓度为接种离体根的干重克数与培养液体积毫升数之比。 [0019] The concentration of inoculum from the root dry weight in grams of body fluid volume and culture than the number of milliliters vaccination.

[0020] 本发明测定植物离体根比生长速率的装置在工作时，是先将该装置按顺序连接到离体根生物反应器上，再把植物离体根接种到生物反应器内，进行培养。 [0020] The present invention was measured in vitro plant root specific growth rate of the device during operation, the device is first connected in order to the body from the root bioreactor, then vitro plant roots inoculated into the bioreactor, carried out bring up. 随着离体根体积的增加，离体根生物反应器里的液体被不定根增长的体积排开到第一刻度管中，当第一刻度管中的液体高度与虹吸管顶端平齐时，根据虹吸原理，第一刻度管与虹吸管中的培养液会全部流入废液瓶中，然后，离体根生物反应器里的液体再随不定根增长排到第一刻度管中。 With the increase in vitro root volume, the volume in vitro root bioreactor liquid is adventitious roots growing into the first row of open graduated tube, when the first calibration tube and siphon the liquid height at the top flush, According to siphon principle, first graduated tube and siphon the broth will all flow into the waste container, and then, in vitro root bioreactor liquid and then discharged with adventitious roots grow first graduated tube. 最终离体根体积增加的量，就等于第二刻度管中的液体指示值加上第一刻度管的液体指示值。 Isolated Root final volume increase of the amount of the calibration tube is equal to the second value plus the liquid in the liquid indicates the instruction value of the first graduated tube. 记录好每天离体根体积增加的量，折算成离体根重量增加的量，最后画出其生长曲线，就可根据公式算出比生长速率。 Record the day vitro root volume increased volume, translated into an increase in the amount of weight in vitro root, and finally draw the growth curve can be calculated according to the formula than the growth rate.

附图说明 Brief Description

[0021] 图1为本发明装置的结构示意图； Structure of the present invention means [0021] Figure 1 a schematic view;

[0022] 图2为实施例中太子参不定根的生长曲线； [0022] Figure 2 shows the growth curve case Taizishen adventitious roots implementation;

[0023] 图3为实施例中太子参不定根的比生长速率。 [0023] FIG. 3 is the specific growth rate case Taizishen adventitious roots implementation.

具体实施方式 DETAILED DESCRIPTION

[0024] 参见图1，本发明中的测定植物离体根比生长速率的装置，包括顺序连接连通的离体根生物反应器6、第一刻度管2、虹吸管7、废液瓶4和第二刻度管3 ;虹吸管7的一端与第一刻度管2的下部连接连通，虹吸管7的另一端插入废液瓶4的底部；第二刻度管3与废液瓶4的下部连接连通。 [0024] Referring to Figure 1, the present invention was measured in vitro plant root specific growth rate, comprising sequentially connected communication excised roots bioreactor 6, a first calibration tube 2, siphon 7, the waste container 4 and The second scale tube 3; siphon end 7 with the first calibration tube connecting the lower 2 connectivity, the other end of the siphon tube into the waste bottle bottom 7 4; 3 and the lower part of the second calibration tube 4 is connected to the waste bottle connectivity. 图中所示，5为植物离体根。 As shown in Fig, 5 for in vitro plant roots.

[0025] 本发明测定植物离体根比生长速率的装置中，第一刻度管2和废液瓶4上均连接有带空气过滤器I的通气管。 [0025] The present invention was measured in vitro plant root specific growth rate means, the first calibration tube 2 and 4 are connected to the waste container with the air filter I snorkel.

[0026] 本发明测定植物离体根比生长速率的装置中，第一刻度管的精度为0.1毫升，其数值反映的是第一刻度管内所收集的液体体积；第二刻度管的精度为I毫升，其数值反映的是废液瓶4内所收集的液体体积。 [0026] Determination of in vitro plants of the present invention, the specific growth rate of the root apparatus, accuracy of the first calibration tube is 0.1 ml, which reflects the value of the first volume of liquid collected in the calibration tube; accuracy of the second of the calibration tube for I ml, its value reflects the volume of liquid waste bottles collected within four.

[0027] 下面以太子参不定根的培养为例进一步说明本发明的测定植物离体根比生长速率的方法。 [0027] The following heterophylla adventitious root culture as an example to further illustrate the body than the growth rate of the root method for the determination of the present invention from the plant.

[0028] 本实施例中，植物离体根体积为10升，应用该发明得出了太子参不定根在24天内的生长曲线如图2所示，比生长率如图3所示。 [0028] In the present embodiment, in vitro plant root volume of 10 liters, the application of the invention shown in Figure 2 obtained heterophylla adventitious roots 24 days in the growth curve shown in FIG. 3 as shown in the specific growth rate.

[0029] 具体的比生长速率如图纵坐标所示，为0. 02-0. 08。 [0029] Specifically specific growth rate as shown in FIG ordinate, 0. 02-0. 08.