Anticancer Activity of Rice Callus Suspension Cultures from Aromatic Varieties and Metabolites Regulated in Treated Cancer Cell Lines

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures(RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

MACS-W:A modified optical clearing agent for imaging 3D cell cultures

Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.

Targeting Cancers: Uncovering the Potential Roles of Potato Tissue Culture as Anti-Cancer Agents - A Secondary Publication

Plant tissue culture is a technique that enhances the quality and quantity of potatoes. Potatoes are a significant crop and are primarily used in the world. It is a staple food in many countries, where millions of tonnes are produced annually. It is an essential source of many nutrients, such as proteins, carbohydrates, vitamins, and beta-carotene. In addition, potatoes are being used as therapeutic agents against cancer and other human diseases as well. Potatoes are on the third list after wheat and rice. To overcome food shortages and malnutrition, there are two methods used for producing potatoes: the first is sexual, which is seed propagation, and the second is asexual, which is plant tissue culture propagation. Conventional potato breeding is a uniform method, but it is unsafe because there is a risk of pathogen attack. In a laboratory setting, the tissue culture of potatoes produced millions of plants with nutrient-rich medium under controlled environmental conditions that prevent pest attacks. Some environmental stresses, such as salinity and water scarcity, affect potato yield and production;however, applying nanoparticles like organic, inorganic, and silicon dioxide enhances potato quality and combats stress. Biotechnology has proven to be helpful in addressing all these issues. This review discusses the significance of potatoes, their production through the tissue culture technique, and the application of nanoparticles to improve the growth, and impact of potatoes on human health.

Lignin Biosynthesis Studies in Plant Tissue Cultures

Lignin, a phenolic polymer abundant in cell walls of certain cell types, has given challenges to scientists studying its structure or biosynthesis. In plants lignified tissues are distributed between other, non-lignified tissues, Characterization of native lignin in the cell wall has been difficult due to the highly cross-linked nature of the wall components. Model systems, like plant tissue cultures with tracheary element differentiation or extracellular lignin formation, have provided useful information related to lignin structure and several aspects of lignin formation. For example, many enzyme activities in the phenylpropanoid pathway have been first identified in tissue cultures. This review focuses on studies where the use of plant tissue cultures has been advantageous in structural and biosynthesis studies of lignin, and discusses the validity of tissue cultures as models for lignin biosynthesis.

Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro

The purpose of this study was to compare cell growth characteristics,ciliated cell differentiation,and function of human nasal epithelial cells established as explant outgrowth cultures or dissociated tissue cultures.Human nasal mucosa of the uncinate process was obtained by endoscopy and epithelial cell cultures were established by explant outgrowth or dissociated tissue culture methods.Epithelial cell growth characteristics were observed by inverted phase contrast microscopy.Ciliated cell differentiation was detected byβ-tubulin IV and ZO-1 immunocytochemistry.Basal and ATP-stimulated ciliary beat frequency(CBF)was measured using a high-speed digital microscopic imaging system.Both the explant and dissociated tissue cultures established as monolayers with tight junctions and differentiated cell composition,with both types of cultures comprising ciliated and non-ciliated epithelial cells.Fibroblasts were also frequently found in explant cultures but rarely seen in dissociated tissue cultures.In both culture systems,the highest ciliated cell density appeared at 7th–10th culture day and declined with time,with the lifespan of ciliated cells ranging from 14 to 21 days.Overall,10%of the cells in explant cultures and 20%of the cells in the dissociated tissue cultures were ciliated.These two cultures demonstrated similar ciliary beat frequency values at baseline(7.78±1.99 Hz and 7.91±2.52 Hz,respectively)and reacted equivalently following stimulation with 100μM ATP.The results of this study indicate that both the explant outgrowth and dissociated tissue culture techniques are suitable for growing well-differentiated nasal ciliated and non-ciliated cells,which have growth characteristics and ciliary activity similar to those of nasal epithelial cells in vivo.

Tissue Culture of Calla Lily (Zantedeschia spreng.): An Updated Review on the Present Scenario and Future Prospects

The calla lily(Zantedeschia spreng.)is a bulbousflower native to the tropical regions of Africa.Calla lily has gained significant popularity in the international market owing to its intricate morphology and prolonged flowering duration.Despite such advantages,for two sub-groups of calla lily,known as group Zantedeschia and group Aestivae,there are challenges in terms of hybrid production due to the‘plastome-genome incompatibility’there-between.Tissue culture is a fundamental biotechnological tool used in gene editing research,with a focus on disease resistance andflower color in calla lily breeding programs.The present review provides a brief background on the history and development of the calla lily,as well as a comprehensive and critical summary of calla lily tissue culture research.The regeneration pathways for both group Zantedeschia and group Aestivae can be divided into de novo organogenesis and somatic embryogenesis.Both groups are capable of obtaining replants through such means.However,only some species in group Aestivae have been reported to be successful in the somatic embryogenesis pathway.In the present review,special attention was paid to the influence of explant types,plant growth regulators,and culture conditions on both de novo organogenesis and somatic embryogenesis in calla lily tissue culture.Ultimately,future research prospects were determined based on integrated analysis of recent progress in calla lily tissue culture research.

Evaluation of Different Substrates Compositions for Acclimatization of Tissue Culture Taro Plantlets in a Propagator

Taro is cultivated in most Regions of Cameroon and it is affected by taro leaf blight disease since 2010 which has decreased its production. Lack of disease-free planting materials has been a main problem to farmers. This study was carried out at International Institute of Tropical Agriculture (IITA) Yaounde and Institute of Agricultural Research for Development (IRAD) Bambui to assess different substrates for acclimatization of tissue culture taro plantlets in apropagator. No information is available on acclimatization of Cameroonian taro plantlets in different substrates. Taro plantlets from tissue culture were acclimatised in a propagator for six weeks under different substrates, the first substrate consisted of sterile three parts of soil and one part of river sand mixed together (3:1), the second substrate consisted of sterile two parts of soil and two parts of river sand mixed together (2:2), the third substrate consisted of sterile two parts of soil, one part of rice husk and one part of river sand mixed together (2:1:1) and the fourth substrate consisted of sterile one part of soil and three parts of river sand mixed together (1:3). After acclimatisation of the different taroplantlets (Dark green petiole with small leaves (L1), Red petiole with small leaves (L2), Light green petiole with large leaves (L3) and Light green petiole with small leaves (L4) in these four substrates, it was observed that the best growth rate of plant was recorded on substrate sand + soil (1:3). The other substrates showed moderate growth of plants. Substrate sand + soil (1:3) can be recommended for acclimatization of Cameroonian taro plantlets.

台湾独蒜兰组培繁育及其假鳞茎的化学组成分析

为构建台湾独蒜兰组培繁育技术体系,分析栽培台湾独蒜兰假鳞茎化学组成特征,经过种子萌发、原球状茎增长、假鳞茎增殖、生根壮苗培养获得台湾独蒜兰植株,以组培台湾独蒜兰植株的假鳞茎为材料,应用非靶向代谢组和HPLC指纹图谱分析假鳞茎化学成分组成。结果表明,经过2年的繁育,单个台湾独蒜兰蒴果可以获得假鳞茎0.5 g左右的幼苗超过1 000株。通过非靶向代谢组数据分析鉴定出不同材料的共有的化合物1 229种,包括羧酸及其衍生物236种、苯及取代衍生物110种、有机氧化合物106种、脂肪酰基化合物94种、黄酮类化合物43种、甾体及甾体衍生物42种,其中以化合物1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯为代表的苄酯类为兰科植物富集化合物。应用HPLC法,测得假鳞茎的1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯含量为0.53%。综上,应用组培技术可以繁育得到台湾独蒜兰的植株,台湾独蒜兰的假鳞茎与独蒜兰一样含有丰富的苄酯类化合物成分。

细胞全能性转录因子调控植物组培再生的分子机制研究进展

植物细胞在适宜的培养条件下展现出发育成完整新个体的潜能,这种潜能被称为植物细胞全能性。基于细胞全能性的组织培养技术,在植物无性繁殖和遗传改良领域得到了广泛应用。非生物胁迫、植物激素与转录因子协同调控植物离体再生过程。其中,在植物再生过程中起主导作用的转录因子,被称为细胞全能性转录因子。近年来,关于细胞全能性转录因子调控植物离体再生的分子机制研究已取得显著进展。众多转录因子被鉴定出来,并在提高植物遗传转化效率的研究中得到了初步应用。本文按照植物细胞全能性转录因子在植物再生中的功能进行分类梳理,综述近几年植物离体再生过程中信号转导机制,总结植物细胞全能性转录因子在提高植物遗传转化效率方面的作用,并对其应用前景进行展望,为建立有效的无性繁殖体系提供科学依据。

“粉玉1号”草莓茎尖组织培养体系及其脱毒效果研究

“粉玉1号”是通过杂交育种方式选育的早熟抗病粉果草莓新品种。从大棚栽培的“粉玉1号”草莓植株上取匍匐茎芽为试料,通过外植体消毒、茎尖剥取、不定芽诱导和生根培养建立了“粉玉1号”茎尖组织培养脱毒技术体系,对茎尖培养苗和大棚栽培植株进行了草莓皱缩病毒(SCV)、草莓斑驳病毒(SMoV)、草莓轻型黄边病毒(SMYEV)和草莓镶脉病毒(SVBV)PCR检测。结果表明,“粉玉1号”草莓外植体灭菌方法为75%乙醇处理30 s,再用0.1%升汞溶液处理10 min;匍匐茎芽一般需剥去1片嫩叶和7片幼叶才能剥出茎尖;茎尖初代培养基宜采用MS+0.5 mg/L 6-BA,继代培养基宜采用MS+0.1 mg/L 6-BA,生根培养基宜采用不加植物生长调节剂的1/2 MS。大棚栽培植株样品存在SVBV感染,通过茎尖组培脱除了该病毒,SCV、SMoV和SMYEV在所有样品中均未检出。建立的茎尖脱毒技术体系可为“粉玉1号”草莓工厂化育苗提供参考。

不同温度调节对云南独蒜兰组培苗生长的影响

通过云南独蒜兰种子无菌萌发获得原球茎;原球茎经过增殖长叶、生根培养获得组培苗;然后在温度为15、20、25、30、35℃下进行分别培养,光照时间12 h/d,光照强度1500 lx。挑选出30℃(T1)和35℃(T2)的组培苗转入变温条件(白天温度25℃、光照时长12 h,晚上温度20℃、光照时长12 h)的培养箱内进行培养,对照(CK)培养条件为温度25℃(光照时长24 h),光照强度均为1500 lx。定期观察云南独蒜兰叶片数量和长度、假鳞茎数量和大小、须根数。结果表明:假鳞茎数量、大小在35℃培养条件下最高,15℃培养条件下最低;叶片长度在25℃培养条件下最高,35℃培养条件下最低;叶片数量在25℃培养条件下最高,35℃培养条件下最低;须根数在35℃培养条件下最高,20℃培养条件下最低。通过变温继续培养后,假鳞茎数量、大小和须根数T2最多,叶片数量和叶片长度CK组最多。通过炼苗对比,传统云南独蒜兰组培苗炼苗成活率为83.3%,成苗率为72.4%,变温处理后云南独蒜兰组培苗炼苗成活率达到94.6%,成苗率达到91.2%。

白城山新1号杨增殖培养试验

为筛选出白城山新1号杨最佳增殖培养基配方,本试验采用正交试验法经组织培养对其进行繁育,以MS培养基为基础配方,将6-BA质量浓度、NAA质量浓度、大量元素含量、蔗糖质量浓度分别设置5个水平进行试验。经回归分析,确定MS+6-BA 0.30 mg·L^(-1)+NAA 0.05 mg·L^(-1)+蔗糖20.00 g·L^(-1)为白城山新1号杨组培增殖培养基优化配方。

7年生马尾松组培苗试验林经营效益

通过收集7年生马尾松组培苗试验林胸径、树高、材积和产脂量数据,利用项目投资的财务效益评估手段对其经济效益进行分析研究,为丰富马尾松人工林高效培育技术体系提供试验依据。结果表明:在当前立地条件和经营措施下,组培苗林分林木胸径、树高、平均材积、蓄积量、出材量为15.5 cm、10.03 m、0.1008 m^(3)、151.20 m^(3)·hm^(-2)、91.5879 m^(3)·hm^(-2),分别比实生苗提高了27.0%、21.9%、89.5%、89.5%、94.9%,中径材林木数量占总株数比例提升了30个百分点,组培苗林分生长优势突出,材种规格获得大幅度提升。采脂后组培苗林分产值、净收益、净现值、内部收益率(IRR)为142869.1元·hm^(-2)、54283.4元·hm^(-2)、19802.4元·hm^(-2)、17.2%,分别比未采脂提高了130.9%、855.1%、331.5%、14.7个百分点,比实生苗林分提高了368.6%、475.8%、203.9%、26.6个百分点。7年生马尾松组培苗林分经济效益显著,在立地条件相似地区应用马尾松组培苗造林,可大幅度提高人工林经营效益。

“以学生为中心”视角下高职院校课程的改革与实践——以“植物组织培养技术”课程为例

高职院校肩负着培养高技能型人才的重任,以立德树人为根本任务,不断深化产教融合,推进职业教育高质量发展。“以学生为中心”的教学理念是推进职业教育高质量发展的具体体现。围绕“以学生为中心”这个主题,从课程思政、教学内容、教学设计与组织、教学评价等方面进行改革与探索,实现教学过程工作化,知识、能力、素质培养同步化,教学效果显著。

马尾松组培苗幼林采脂试验分析

【目的】研究马尾松组培苗幼林产脂量与生长性状间的变化规律和松脂主要组分及含量特征,为马尾松早期利用和高产优质脂用原料林培育提供试验依据。【方法】以7年生马尾松组培苗人工林为研究对象,设置标准地开展采脂试验,测定胸径和树高,计算单株材积和产脂量,进行不同径级(树高级)间的产脂量比较分析,并拟合产脂量与径级和树高方程;通过气相色谱/质谱联用技术(GC/MS)检测松脂样品主要化学组分和含量。【结果】7年生马尾松组培苗幼林平均产脂量为15.9 g·d^(-1)·株^(-1)。不同径级产脂量间差异显著(P<0.05),随林木径级增大产脂量随之增加,呈一元线性关系(R^(2)=0.99);不同树高级产脂量随树高增大而增加,呈幂函数关系(R^(2)=0.98)。胸径、树高和材积与产脂量间相关性极显著(P<0.01),可以实现同步改良。7年生马尾松组培苗幼林单位面积可采脂林木数量和产脂量分别是同等条件下实生苗林分的1.9倍和1.8倍。在40%的强度下,采脂对林木生长的影响尚未表现出明显差异。兼顾产脂量和生长性状,马尾松组培苗幼林起始采脂径级以18 cm径级为宜。从组培苗林分松脂样品中共检出α-蒎烯、β-蒎烯、长叶烯、石竹烯、海松酸、山达海松酸、枞酸等单萜、倍半萜和二萜类化学物质22种,其主要组分及含量与常规经营中龄以上马尾松林分松脂基本一致。【结论】该立地条件下营造的7年生马尾松组培苗幼林产脂量、松脂品质已基本满足行业需求,在规范采脂的前提下可对其进行生产性采脂利用。采用马尾松组培苗造林在提早采脂和增加单位面积松脂产量方面具有较大的优势和潜力,为实现马尾松的早期化利用和松脂原料林的高质量培育提供了新的选择。

珠芽魔芋组织诱导快速繁殖技术研究

以珠芽魔芋球茎为外植体,开展组织诱导快速繁殖技术研究,以期为珠芽魔芋的规模化种植提供技术支撑。结果表明:最佳萌发培养基为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+30 g/L蔗糖+5 g/L琼脂,萌发率达100%;最佳愈伤组织诱导培养基为MS+2.0 mg/L 6-BA+0.5 mg/L GA3+30 g/L蔗糖+5 g/L琼脂,叶片和叶柄形成愈伤组织的诱导率分别为99.17%、92.50%;最佳不定芽诱导培养基为MS+1.0 mg/L 6-BA+0.4 mg/L NAA+25 g/L蔗糖+5 g/L琼脂,叶片和叶柄不定芽诱导率分别为94.17%、65.83%,单位愈伤组织分化芽数分别为3、1.87个;最佳生根培养基为1/2 MS+0.4 mg/L NAA+0.2 mg/L IBA+0.5 g/L碳粉+20 g/L蔗糖+5 g/L琼脂,生根率为97.50%,平均生根数为3.0条,平均根长为3.0 cm;炼苗10 d后,移栽到配比为表土∶椰砖营养土∶黑沙土=1∶2∶1的混合基质中,移栽成活率达到91.7%。

木兰科植物组织培养技术研究进展

对木兰科植物的组织培养技术研究现状进行了概述,包括外植体的选择、培养基的选择、继代增殖培养、生根培养及体细胞胚的诱导5个方面,并分析了木兰科植物组培过程中遇到的难题及未来努力的方向,为木兰科植物利用组培快繁技术来获取优质种苗提供参考。

不同叶面肥及株型控制对中科猕砧2号猕猴桃组培苗生长发育的影响

以中科猕砧2号猕猴桃2月龄组培苗为研究对象,研究不同配比的叶面施肥结合株型控制对植株干径生长、叶绿素含量、氮磷钾含量以及幼苗生理指标的影响。结果表明,中科猕砧2号组培穴盘苗的干径、叶绿素含量及植株全氮、全钾含量,根冠比在不同处理间均存在显著差异,植株全磷含量在各处理间差异不显著。4个控制因子中,株型控制对中科猕砧2号组培穴盘苗生长的促进作用最大,全抹芽的株型控制+施复合叶面肥(0.2%尿素+0.1%磷酸二氢钾+0.2%园艺肥)为中科猕砧2号组培穴盘苗育苗期间的最优处理配方。

药用植物组织培养学创新教学模式改革研究

药用植物组织培养学是中草药栽培与鉴定专业的专业基础课,对学生的专业基础理论学习与实践应用能力的培养具有重要意义,是现代植物学与生理学的基础学科。基于我国教育体系的改革进步,以及对应用型人才的培养要求,线下授课已经不能完全满足对学生的授课需求,本文分析了目前教学中存在的几点问题,融合思政教学内容,对教学方法进行创新和改革,以取得良好的教学效果。

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