The objective of this study was to set up a plant micropropagation facility to mass propagate sugarcane, energy cane, and related clonally propagated species. An efficient methodology for micropropagation of energy cane and perennial grasses using temporary immersion bioreactors was developed. Several different methods of tissue culture initiation, multiplication, and rooting were evaluated for several varieties of sugarcane (Saccharum officinarum L.) and sugarcane-related species such as Erianthus spp., Miscanthus spp., and Sorghum spp. × sugarcane hybrids, all from a germplasm collection. Apical meristem cultures were initiated for all genotypes that were micropropagated, when liquid or semisolid Murashige and Skoog (MS) medium was used, which was supplemented with 0.1–0.2 mg L⁻¹ BAP, 0.1 mg L⁻¹ kinetin, 0–0.1 mg L⁻¹ NAA, and 0–0.2 μg L⁻¹ giberellic acid. These cultures produced shoots between 4 and 8 wk after initiation. Shoot regeneration from leaf rolls or immature inflorescences was observed as early as 4 wk after initiation. Shoot multiplication was successful for all genotypes cultured in MS medium with 0.2 mg L⁻¹ BAP and 0.1 mg L⁻¹ kinetin. Energy cane had a significantly higher combined multiplication rate when grown under four or five LED lamps than when grown under three LED lamps, or under fluorescent lights in a growth chamber. The addition of 2 mg L⁻¹ NAA produced faster and better rooting in all of the genotypes tested. Shoots produced well-developed roots after one cycle of 15–21 d in the bioreactors. The maximum number of plantlets produced per bioreactor was 1080. Plantlets developed a vigorous root system and were ready to be transplanted into the field after 2 mo. A protocol was standardized for different energy cane clones that were recommended for their biomass production and cell wall composition. Different tissues were used to speed up or facilitate tissue culture initiation. Visual assessment of micropropagated plants in the field did not show any off-types, based on gross morphological changes of plant morphology or disease reaction, compared to plants of the same genotype derived from a traditional propagation method (stem cuttings). This is the first report of energy cane and Miscanthus spp. micropropagation using the SETIS bioreactor.

这项研究的目的是建立一种植物微繁殖设施，以大规模繁殖甘蔗，能量棒和相关的无性繁殖物种。开发了一种使用临时浸入式生物反应器对甘蔗和多年生禾草进行微繁殖的有效方法。对几种不同的甘蔗（Saccharum officinarum L.）和与甘蔗有关的物种（如Erianthus spp。，Miscanthus spp。和Sorghum）的组织培养起始，繁殖和生根的几种不同方法进行了评估。spp。×甘蔗杂种，均来自种质资源。当使用液体或半固体Murashige和Skoog（MS）培养基，并补充了0.1–0.2 mg L ^-1 BAP，0.1 mg L ^-1激动素，0–0.1 mg的液体或半固体Murashige和Skoog培养基时，针对所有微繁殖的基因型启动了顶峰分生组织培养。L ^-1 NAA和0-0.2μgL ^-1吉贝酸。这些培养物在起始后4至8周之间产生芽。始于4周时，观察到从卷叶或未成熟花序再生的嫩芽。对于在含有0.2 mg L ^-1 BAP和0.1 mg L ^-1的MS培养基中培养的所有基因型，芽繁殖均成功激动素。在四个或五个LED灯下生长时，与在三个LED灯下或在生长室内的荧光灯下生长时相比，能量棒具有显着更高的组合繁殖率。2 mg L ^-1的添加NAA在所有测试的基因型中产生更快更好的生根。在生物反应器中经过15-21 d的循环后，新芽产生了发育良好的根。每个生物反应器最多可生产1080个小植株。小植株发育出旺盛的根系，并准备在2个月后移植到田间。针对不同的能量甘蔗克隆，针对其生物量生产和细胞壁组成推荐了协议。使用不同的组织来加速或促进组织培养的开始。与从传统繁殖方法（干插条）衍生的相同基因型的植物相比，基于植物形态或病害反应的总体形态变化，对田间微繁植物的目视评估未发现任何异型。这是能源甘蔗和芒草属。使用SETIS生物反应器进行微繁殖。