Name: employing aeroponic systems for the clonal propagation of cannabis
Uploaded: 2021-12-01T13:00:00
Description: Watch this Scientific Journal Video about Employing Aeroponic Systems for the Clonal Propagation of Cannabis at JoVE.com

这项研究得到了科罗拉多州立大学普韦布洛分校大麻研究所和科学与信息通信技术部（2021-DD-UP-0379）和春川市（大麻研发和工业化，2020-2021）的支持，作者还希望感谢Summit CBD的Justin Henderson对"樱桃酒"种子的慷慨捐赠。

1. 生成用于克隆繁殖的母株
<ol><li>选择一种健康的雌性母株，该母株表现出与其预期用途特有的理想形态和化学特征。</li>
	<li>让母株达到适当的大小（大约25个成熟芽）以进行克隆繁殖（即插条）。</li>
	<li>让母株保持在营养生长阶段（浅色：深色= 18小时：6小时），以促进芽生长以备将来繁殖。</li>
</ol>2. 气培系统的建设和制备
<ol><li>首先将盖子放?...

<ol>
	<li>ElSohly, M. A., Radwan, M. M., Gul, W., Chandra, S., Galal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phytochemistry+of+Cannabis+sativa+L.">Phytochemistry of Cannabis sativa L.</a> Progress in the Chemistry of Organic Natural Products. 103, 1-36 (2017).</li><li>Cunetti, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+pain+treatment+with+cannabidiol+in+kidney+transplant+patients+in+Uruguay.">Chronic pain treatment with cannabidiol in kidney transplant patients in Uruguay.</a> Transplantation Proceedings. 50 (2), 461-464 (2018).</li><li>Hausman-Kedem, M., Menascu, S., Kramer, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+CBD-enriched+medical+cannabis+for+treatment+of+refractory+epilepsy+in+children+and+adolescents+-+An+observational,+longitudinal+study.">Efficacy of CBD-enriched medical cannabis for treatment of refractory epilepsy in children and adolescents - An observational, longitudinal study.</a> Brain &amp; Development. 40 (7), 544-551 (2018).</li><li>Linge, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cannabidiol+induces+rapid-acting+antidepressant-like+effects+and+enhances+cortical+5-HT/glutamate+neurotransmission:+role+of+5-HT1A+receptors.">Cannabidiol induces rapid-acting antidepressant-like effects and enhances cortical 5-HT/glutamate neurotransmission: role of 5-HT1A receptors.</a> Neuropharmacology. 103, 16-26 (2016).</li><li>Lehmann, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+cannabidiol+treatment+reduces+early+pancreatic+inflammation+in+type+1+diabetes.">Experimental cannabidiol treatment reduces early pancreatic inflammation in type 1 diabetes.</a> Clinical Hemorheology and Microcirculation. 64 (4), 655-662 (2016).</li><li>Shannon, S., Lewis, N., Lee, H., Hughes, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cannabidiol+in+anxiety+and+sleep:+A+large+case+series.">Cannabidiol in anxiety and sleep: A large case series.</a> The Permanente Journal. 23, 18-41 (2019).</li><li>Russo, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=History+of+cannabis+and+its+preparations+in+saga,+science,+and+sobriquet.">History of cannabis and its preparations in saga, science, and sobriquet.</a> Chemistry &amp; Biodiversity. 4 (8), 1614-1648 (2007).</li><li>Vera, C. L., Hanks, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemp+production+in+Western+Canada.">Hemp production in Western Canada.</a> Journal of Industrial Hemp. 9 (2), 79-86 (2004).</li><li>. Hot hemp: How high THC levels can ruin a legal hemp harvest Available from: <a target="_blank" href="https://www.westword.com/marijuana/hot-hemp-how-high-thc-levels-can-ruin-a-legal-hemp-harvest-9963683">https://www.westword.com/marijuana/hot-hemp-how-high-thc-levels-can-ruin-a-legal-hemp-harvest-9963683</a> (2018)</li><li>Lata, H., Chandra, S., Techen, N., Khan, I. A., ElSohly, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+the+genetic+stability+of+micropropagated+plants+of+Cannabis+sativa+by+ISSR+markers.">Assessment of the genetic stability of micropropagated plants of Cannabis sativa by ISSR markers.</a> Planta Medica. 76 (1), 97-100 (2010).</li><li>Caplan, D., Dixon, M., Zheng, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+rate+of+organic+fertilizer+during+the+flowering+stage+for+Cannabis+grown+in+two+coir-based+substrates.">Optimal rate of organic fertilizer during the flowering stage for Cannabis grown in two coir-based substrates.</a> HortScience. 52 (12), 1796 (2017).</li><li>Monthony, A. S., Page, S. R., Hesami, M., Jones, A. M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+past,+present+and+future+of+Cannabis+sativa+tissue+culture.">The past, present and future of Cannabis sativa tissue culture.</a> Plants (Basel). 10 (1), 185 (2021).</li><li>Clarke, R. C., Merlin, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cannabis+domestication,+breeding+history,+present-day+genetic+diversity,+and+future+prospects.">Cannabis domestication, breeding history, present-day genetic diversity, and future prospects.</a> Critical Reviews in Plant Sciences. 35 (5-6), 293-327 (2016).</li></ol>

随着对大麻素含量一致的 大麻 植物的需求不断增加 ，大麻工业 中已经利用了各种克隆繁殖方法。与有性方法相比，无性繁殖显示出大规模，一致生产的几个优点。气培繁殖系统是水培系统的改良版本，利用充气的营养丰富的水雾来提供快速的根部发育。所描述的气培系统由三个关键步骤组成，1）产生健康的雌性 大麻 "母"植物作为所需物理/化学性状的遗传源，2）切割芽（?...

大麻 L.是一种一年生的，雌雄异株的开花植物，属于大麻科。大麻素主要在位于雌性花序苞片组织外表皮层的腺体毛状体内产生1，正在成为越来越受欢迎的研究课题，主要是因为它们逐渐认识到药用特性。大麻二酚（CBD）是 大麻 中仅次于Δ9-四氢大麻酚（THC）的第二大最突出的大麻素，具有许多药用益处，包括镇痛特性2，抗癫痫特性3，抗抑郁特性4，降低糖尿病风险5和治疗各种睡眠障碍6。由于 大麻 植物的代谢物具有多种健康益处，因此对其商业规模生产的需求不断增长7。为了满足这一需求，种植方法不断得到改进和重塑，以不断为新兴的 大麻 行业提供一致，高质量的植物材料。
大麻的繁殖可以通过两种方式促进：有性繁殖或无性繁殖。有性繁殖的一个例子是用雄性雄蕊的花粉为雌性胚珠授粉，从而产生可以发芽的种子。种子发芽是一种可靠的栽培方法，已用于育种和栽培目的，其中在亲本系中选择理想的表型性状，以提高后代大麻植物的质量，包括耐旱性状，抗虫性状，增加产量和增加效力等性状8.然而，在进行有性繁殖时，意外的异花授粉是一种固有的风险，导致不良后代，从而导致理想性状的潜在丧失或引入不需要的性状。大麻种植者接受用THC生产花粉授粉的大麻种子，导致由于不合规的植物（>0.3%总THC w / w）而导致的重大经济损失，突出了这种意外授粉的一个例子9。此外，为了生产仅由雌性组成的作物，必须播种女性化的种子而不是非女性化的种子，这可能导致雌雄同体和其他不良特征，从而导致经济损失。为了克服大麻有性繁殖的局限性，无性繁殖已在大麻行业的商业生产模式中广泛实施10。
大麻的无性繁殖只需要一种植物，这允许单一基因型的繁殖，从而允许商业生产具有理想农艺和药物性状的植物。无性大麻繁殖的一种常见形式是将雌性植物的一小部分切割并插入无土基质11中，该基质被湿度圆顶覆盖以诱导根的形成。虽然这种方法已被证明是成功的，但一个共同的缺点是圆顶内积聚了高水平的湿度（通常为80%或更高），为真菌病原体提供了理想的生长环境，这可能对新的敏感插条有害。另一种形式的无性繁殖是使用组织培养的微繁殖，其中无菌技术允许昆虫，微生物和无病毒的大麻植物材料在有限的空间内繁殖12。然而，这个过程是昂贵，耗时的，需要训练有素的实验室技术人员，而大型大麻设施通常无法进入。
关于 大麻克隆繁殖的已发表研究报告很少。为了为理解用于研究目的和工业生产的 大麻 的无性繁殖提供基础，本研究旨在证明采用气培系统进行 大麻克隆繁殖的易用性和可及性。气培系统是 大麻无性繁殖的理想选择，始终如一地为插条提供营养丰富的水，及时诱导早期根系形成，并在需要时允许植物无限期地维持。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1-part Fox Farm</td>
			<td>Fox Farm</td>
			<td></td>
			<td>Soil Mix</td>
		</tr>
		<tr>
			<td>1-part Promix</td>
			<td>Promix</td>
			<td></td>
			<td>Soil Mix</td>
		</tr>
		<tr>
			<td>1-part Roots Organic Original</td>
			<td>Auora Innovations</td>
			<td></td>
			<td>Soil Mix</td>
		</tr>
		<tr>
			<td>1-part Wiggle Worm Earth Worm Castings</td>
			<td>UNCO Industries</td>
			<td></td>
			<td>Soil Mix</td>
		</tr>
		<tr>
			<td>Algae and Bacterial Cleaning Solution (Clear Rez)</td>
			<td>EZ Clone</td>
			<td>SKU#: 225</td>
			<td>8 fl. Oz.</td>
		</tr>
		<tr>
			<td>Artificial Lighting</td>
			<td>AgroBrite</td>
			<td>SKU#: 1399</td>
			<td>T5 324W 4&#39; 6-Tube Fixture with Lamps</td>
		</tr>
		<tr>
			<td>Cannabis Mother plant 1 (Cherry Wine)</td>
			<td>Summit CBD</td>
			<td>N/A</td>
			<td>Donated material</td>
		</tr>
		<tr>
			<td>Cannabis Mother Plant 2 (Red Wine)</td>
			<td>Trilogene</td>
			<td>SKU: 0101RR</td>
			<td></td>
		</tr>
		<tr>
			<td>Corresponding Plastic Lid</td>
			<td>Office Depot</td>
			<td>N/A</td>
			<td>38.1 cm x 25.4 cm</td>
		</tr>
		<tr>
			<td>Drill Bit 1</td>
			<td>Dewalt</td>
			<td>DW1586</td>
			<td>38.1 mm spade drill bit</td>
		</tr>
		<tr>
			<td>Drill Bit 2</td>
			<td>Dewalt</td>
			<td>DW1308</td>
			<td>3.175 mm drill bit</td>
		</tr>
		<tr>
			<td>Flora/Bloom (Nutrient Solution)-5 mL</td>
			<td>General Hydroponics</td>
			<td>SKU#: 726</td>
			<td>946 mL (1 Quart) 2.43 lbs. (1.1 kg) (Available Phosphate 5.0%, Soluble Potash 4.0%, Magnesium 1.5%, Sulfur 1.0%)</td>
		</tr>
		<tr>
			<td>FloraGrow (Nutrient Solution)- 5 mL</td>
			<td>General Hydroponics</td>
			<td>SKU#: 724</td>
			<td>946 mL (1 Quart) 2.43 lbs. (1.1 kg) ((Total Nitrogen 2.0% (0.25% Ammoniacal Nitrogen, 1.75% Nitrate Nitrogen), Available Phosphate 1.0%, Soluble Potash 6.0%, Magnesium 0.5%))</td>
		</tr>
		<tr>
			<td>FloraMicro (Nutrient Solution)- 5 mL</td>
			<td>General Hydroponics</td>
			<td>SKU#: 759</td>
			<td>946 mL (1 Quart) 2.43 lbs. (1.1 kg) ((Total Nitrogen 5.0% (0.3% Ammoniacal Nitrogen, 4.7% Nitrate Nitrogen), Soluble Potash 1.0%, Calcium 5.0%, Boron 0.01%, Cobalt 0.0005%, Copper 0.01%, Iron 0.1%, Manganese 0.05%, Molybdenum 0.0008%, Zinc 0.015%))</td>
		</tr>
		<tr>
			<td>Horticultural Scissors</td>
			<td>Shear Perfection</td>
			<td>SKU#: 12620</td>
			<td>Platinum Stainless Steel Bonsai Scissors (2.4&#34;)</td>
		</tr>
		<tr>
			<td>Isopropyl Alcohol</td>
			<td>Equate</td>
			<td>Walmart # 574133562</td>
			<td>70% concentration</td>
		</tr>
		<tr>
			<td>Nutrient Mist Solution (Clonex Mist)</td>
			<td>Growth Technology</td>
			<td>SKU#: 4889</td>
			<td>10.14 fl. Oz (300 ml) (Total Nitrogen: 5.9 &#215; 10-4 %, Available Phosphate: 4.0 &#215; 10-4 %, Soluble Potash: 5.0 &#215; 10-4 %)</td>
		</tr>
		<tr>
			<td>pH Down</td>
			<td>General Hydroponics</td>
			<td>SKU#: 733</td>
			<td>946 ml (1 Quart) 2.43 lbs. (1.1 kg)</td>
		</tr>
		<tr>
			<td>pH Up</td>
			<td>General Hydroponics</td>
			<td>SKU#: 730</td>
			<td>946 ml (1 Quart) 2.43 lbs. (1.1 kg)</td>
		</tr>
		<tr>
			<td>Plastic Container</td>
			<td>Office Depot</td>
			<td>N/A</td>
			<td>38.1 cm x 25.4 cm x 30.48 cm</td>
		</tr>
		<tr>
			<td>Power Drill</td>
			<td>Dewalt</td>
			<td>DCD709B</td>
			<td>20-Volt Max &#189;&#8221; Drill</td>
		</tr>
		<tr>
			<td>Rockwool Cubes</td>
			<td>Grodan</td>
			<td>SKU#: 830</td>
			<td>38.1 mm</td>
		</tr>
		<tr>
			<td>Rooting Solution (Clonex Rooting Gel)</td>
			<td>Growth Technology</td>
			<td>SKU#: 939</td>
			<td>3.4 fl. Oz. (100 ml) (Indolebutyric Acid - 0.31%)</td>
		</tr>
		<tr>
			<td>Statistic Software (Prism)</td>
			<td>GraphPad Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Submersible Water Pump</td>
			<td>ActiveAQUA</td>
			<td>SKU: AAPW250</td>
			<td>Model: AAPW250, Voltage 120V, Power 16W</td>
		</tr>
	</tbody>
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employing aeroponic systems for the clonal propagation of cannabis

该协议描述了通过利用气培系统实现大麻有效克隆繁殖技术的标准化。从两个大麻品种中切除了主要的芽插条，命名为"樱桃酒"和"红罗宾"（17-20%w / w CBD），作为"母株"。在放入系统之前，应用生长素前体（吲哚-3-丁酸）来刺激切除插条基部的根系发育。插条每三天用营养雾溶液轻轻喷雾，以提供营养支持，因为溶液含有必需的常量营养素，包括氮，磷和钾。气培系统储水器的pH值范围保持在5.0-6.0之间，水温在20-22°C之间。 使用潜水水泵将水输送到岩屑中。每天为芽尖插条提供24小时的光照，持续10天，直到根部发育发生，在此上移植根部插条用于研究目的。这些气培系统已被证明可以为 大麻 的繁殖产生理想的结果。这里描述的方法减轻了传统方法产生的潜在时间限制，以便为 大麻的无性繁殖提供更有效的手段。

为了验证所描述的气培系统的效率，分别从母株中切除了10个和12个健康的14厘米长的芽，分别是"樱桃酒"和"红罗宾"（图1A，B）。浸入生根诱导培养基后，将克隆放入系统（图2A）。气培系统的构造和运行如图 2A所示。
适应2天后，所有克隆在3-7天内开始发育根系，并在系统上10-14天后完全发育出37 ...

Watch this Scientific Journal Video about Employing Aeroponic Systems for the Clonal Propagation of Cannabis at JoVE.com

Employing Aeroponic Systems for the Clonal Propagation of Cannabis

该协议旨在通过实施气培系统为 大麻苜蓿 L.的克隆繁殖提供指导信息。这里描述的方法包括成功再现大麻属中理想的形态学和化学性质所需的所有用品和 方案。

采用气培系统进行大麻的克隆繁殖

This protocol describes the standardization of an efficient clonal propagation technique of hemp by utilizing aeroponic systems. Primary shoot cuttings ...

该协议可以帮助就如何在研究或商业生产环境中继续进行所需大麻遗传学的再生做出明智的决定。该技术的主要优点是诱导大麻插条中的早期根形成，同时限制其对潜在病原体的易感性。该技术的意义延伸到通过诱导插条中的早期根形成来改善在大麻繁殖中观察到的挑战，这可以应用于研究或生产环境。用无菌的手术刀或剪刀，在顶端分生组织附近以45度角切下具有几个长约10厘米的节点的芽。除前三个节点上存在的树叶外，请删除所有树叶。将新切除的切口从茎基部向上约2至5厘米浸入含有吲哚-3-丁酸的生根溶液中约五秒钟。将切口插入岩棉立方体的中心，直到立方体底部上方约两厘米的深度，位于气培系统中。每三天用营养喷雾溶液喷洒无根的插条。每天在光照下生长插条约18至24小时，光合光子通量密度为每平方米每秒100微摩尔。每两到五天，用pH值在五到六之间的水补充系统。每三天用营养雾溶液轻轻喷洒插条，每三到五天将五毫升营养液倒入储液槽。每五天加入15毫升藻类和细菌清洁溶液，含有0.028%次氯酸。选择具有长而白色纤维根的插条。小心地将岩棉立方体从系统中移开并解开根部。将大麻繁殖体移植到一个装满营养土壤混合物的四升育婴盆中。该图显示了在三到七天内长出根的克隆，并在系统上10到14天后完全发育出37厘米长的根，种植在充满土壤的花盆中。樱桃酒的芽和根的平均长度分别约为24.8厘米和37.8厘米，红知更鸟的平均长度约为21.4厘米和39.7厘米。采用双向方差分析分析两个品种之间的差异，然后进行Tukey多重同情试验，显示两个品种的芽长和根长无显著差异。选择表现出旺盛新生长的健康插条是繁殖的理想选择。保持系统储液槽的水位、温度、pH 值和营养方面是快速有效结果的关键。

Research

JoVE Journal

Bioengineering

Biomolecular Detection employing the Interferometric Reflectance Imaging Sensor (IRIS)

Biomolecular Detection employing the Interferometric Reflectance Imaging Sensor IRIS

生物分子检测的干涉反射成像传感器（IRIS）

The sensitive measurement of biomolecular interactions has use in many fields and industries such as basic biology and microbiology, ...

科研

生物工程

Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems

在生物系统中的离子浓度测量荧光纳米粒子

Tightly regulated ion homeostasis throughout the body is necessary for the prevention of such debilitating states as dehydration.1 In contrast, rapid ion ...

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

海藻酸钠微胶囊作为一个人类胚胎干细胞（胚胎干细胞）的繁殖和分化的3D平台，以不同的谱系

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

Magnetic Resonance Elastography Methodology for the Evaluation of Tissue Engineered Construct Growth

磁共振弹性成像方法评价组织工程构造增长

Traditional mechanical testing often results in the destruction of the sample, and in the case of long term tissue engineered construct studies, the use ...

Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion

三维细胞培养模式的测量肿瘤细胞浸润间质流体流动的影响

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling ...

Continuous High-resolution Microscopic Observation of Replicative Aging in Budding Yeast

在芽殖酵母连续高分辨率显微观察复制性衰老

We demonstrate the use of a simple microfluidic setup, in which single budding yeast cells can be tracked throughout their entire lifespan. The ...

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

在光刻技术定义的聚对二甲苯-C细胞图案：二氧化硅<sub&gt; 2</sub&gt;基板

Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain ...

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy

调查纤维素酶与基于AFM的单分子力谱受体 - 配体系统

Cellulosomes are discrete multienzyme complexes used by a subset of anaerobic bacteria and fungi to digest lignocellulosic substrates. Assembly of the ...

Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study

烟草中瞬时表达的蛋白作为一个案例研究：复杂系统采用的实验方法设计的表征

Plants provide multiple benefits for the production of biopharmaceuticals including low costs, scalability, and safety. Transient expression offers the ...

Merging Ion Concentration Polarization between Juxtaposed Ion Exchange Membranes to Block the Propagation of the Polarization Zone

并列的合并离子交换膜之间的离子浓差极化，以块极化区的传播

The ion concentration polarization (ICP) phenomenon is one of the most prevailing methods to preconcentrate low-abundance biological samples. The ICP ...