Morphogenic Response of Leaf and Petiole Explants of Broccoli Using Thidiazuron

Broccoli (Brassica oleracea var. italica) is an important nutritionally rich vegetable cole crop grown in the world. Environmental stress, pests, and diseases cause enormous yield losses because of a limited gene pool. Genetic manipulation is becoming an important method for broccoli improvement. The objective of present study was to evaluate the potency of thidiazuron (TDZ) as a plant growth regulator in evoking morphogenic responses in leaf and petiole explants of broccoli. An efficient, reproducible, and high frequency plant regeneration protocol has been standardized in broccoli cv. Solan green head. Leaf and petiole explants were cultured on Murashige-Skoog (MS) medium, supplemented with a wide range of TDZ concentrations. The following treatments were designed for efficient in vitro shoot regeneration: TDZ alone, TDZ with adenine, TDZ with naphthalene acetic acid (NAA), and TDZ with indole acetic acid (IAA). Among the 36 combinations of growth regulators used, the highest percentage of leaf explants producing shoot (89.25%) was recorded on MS medium containing 1.0 μM TDZ and 0.107 μM NAA. The multiple shoot regeneration response of petiole explant producing shoots (91.55%) was obtained on MS medium containing 2.0 μM TDZ and 0.107 μM NAA. Shoot multiplication and elongation were obtained on the same medium. For root regeneration in in vitro regenerated shoots, different concentrations of NAA were applied. High frequency (100%) root regeneration response with healthy and vigorous roots was observed on MS medium supplemented with 0.54 μM NAA. The regenerated plantlets with well-developed shoots and root system were transferred to pots containing cocopeat and successfully acclimatized. We recommend 1.0 μM TDZ with 0.107 μM NAA and 2.0 μM TDZ and 0.107 μM NAA combinations for adventitious shoot regeneration from leaf and petiole explants in broccoli cv. Solan green head respectively. This is the first report on high frequency organogenesis from leaf and petiole explants of broccoli cv. Solan green head using thidiazuron.     

小麦茎生长点转化研究初报

为了躲避转基因技术对组织培养的过分依赖,本研究以小麦为材料尝试了一种对生长点直接进行转化的方法,并初步证明了这一方法的可行性。具体做法是：将种子萌芽,然后剥去胚芽鞘暴露出生长点,再用玻璃纤维制作的小刷子将生长点刺伤,最后用带有外源基因的农杆菌进行侵染处理。用携带hph-GUS基因的LBA4404农杆菌,侵染处理1～11日龄幼苗(生长点),共培养7d后检测新生芽中GUS的瞬时表达情况;侵染1～2日龄幼苗(生长点),取长成植株的2～3朵小花进行GUS稳定表达检测。又用携带BADH和npt Ⅱ基因的AGL1农杆菌菌株侵染1日龄的幼苗(生长点),在含：100mg／L卡那霉素的蛭石中进行选择。结果表明,GUS的瞬时表达率随被处理幼苗的日龄增加而降低,以2日龄幼苗为最高(10．7％)。用花序检测GUS的稳定表达,被侵染受体为1日龄幼苗时的表达率高于2日龄的幼苗。蔗糖的存在并不提高GUS基因在花序中表达的频率。不过花序表现为GUS阳性的植株后代,经PCR检测后并没有证实GUS基因存在。用AGL1菌株进行转化,获得了13．6％的抗卡那霉素的绿色植株,但只有3株呈现PCR阳性,且只有1株结了实。     

An efficient in vitro shoot regeneration through direct organogenesis from seedling-derived petiole and leaf segments and acclimatization of Ficus religiosa

Prolific and rapid in vitro plant organogenesis via direct regeneration has been obtained from axenic seedling-derived petiole and leaf explants of Ficus religiosa in Murashige and Skoog (MS) medium containing different concentrations of cytokinins in combination with indole-3-butyric acid (IBA). MS medium with 1.5 mg/l 6-benzylaminopurine plus 0.15 mg/l IBA produced the highest shoot induction frequency with an average of 6.26 and 10.13 shoots per leaf and petiole explants, respectively. After 4 weeks, the highest root formation frequency (96.7%), root number (5.73), and root length (4.76 cm) were with MS medium containing 2.0 mg/l IBA plus 0.1 mg/l α-naphthalene acetic acid. In addition, the effect of four sodium nitroprusside (SNP) treatments on acclimatization was also studied. Highest morphological traits such as survival rates, fresh and dry root weights as well as antioxidant enzymatic activities such as superoxide dismutase, peroxidase, and catalase was achieved with 125 ppm SNP. The α-amino acid, proline, content was highest with this treatment while the highest H2O2 (hydrogen peroxide) was in the controls. This study introduces a cost-effective, prolific, and efficient in vitro multiplication system to supply pharmaceutical and ornamental needs. It is the first report of an in vitro organogenesis protocol for F. religiosa by direct regeneration through axenic seedling-derived petiole and leaf explants, which can be efficiently employed for the utilization of active biomolecules.     

茬次和一天内不同收获时间对紫花苜蓿青贮发酵质量和体外发酵参数的影响

为了研究茬次和一天内不同收获时间对紫花苜蓿青贮饲料品质的影响,并进一步评价其饲用价值,本试验采用双因素设计,两茬紫花苜蓿于现蕾期同一天08：00（AM）,13：00（M）和18：00（PM）刈割并调制青贮饲料,室温下保存45 d后,取样分析并采用体外发酵法评价其饲用价值。结果表明,第1茬紫花苜蓿青贮饲料的相对饲喂价值（relative feed value,RFV）,干物质体外消化率（in vitro dry matter digestibility,IVDMD）及体外发酵挥发性脂肪酸（volatile fatty acids,VFA）、产气量均显著高于第2茬（P<0.05）。下午收获的紫花苜蓿青贮饲料有更高的干物质（dry matter,DM）含量及IVDMD （P<0.05）。因此,在实践生产中,要抓好第1茬的苜蓿生产,推迟刈割时间至中午或下午,可获得更高品质的紫花苜蓿青贮饲料。     

A simple method to obtain endophytic microorganisms from field-collected roots

We describe a simple technique for obtaining endophytic microorganisms from field-collected roots. Roots of plant species were surfaced-sterilized, cut into pieces, and then each root piece was transferred to drops of Gel-Gro medium. The number of segments with bacteria and fungi was quantified and those with bacteria and dark septate endophytic (DSE) fungi were plated onto tryptic soy agar (TSA) and malt extract agar (MEA), respectively. When arbuscular mycorrhizal fungi (AMF) grew from root segments, they were inoculated into Ri T-DNA-transformed carrot root cultures. Eight different species of bacteria and five dark septate endophytes were isolated. Thirty AMF isolates were identified and continuous pure cultures established. This easy and inexpensive approach allowed us to culture various endophytic microorganisms in an in vitro system saving time and space.     

2022年凤阳县水稻品种展示试验初报

凤阳县近几年种子市场销售的杂交水稻品种众多,农民选种是个难题。农业部门为了筛选出适宜当地大面积推广的高产、优质、广适的水稻品种,2022年征集了全县种植面积较大的35个品种参加展示试验。通过专人负责田间管理、田间观察记载、室内考种、总结等,得出结论：徽两优166、Y两优1577、N两优1133、晶两优1377、Q两优851、诺两优6号、天龙两优140、韵两优332、Y两优808、徽两优粤农丝苗等10个品种的抗逆性、丰产性、适应性较好,可以在凤阳县大面积种植推广。     

植物叶片葡萄糖活体检测双通道传感器的制备及应用

【目的】制备植物叶片葡萄糖活体检测传感器,以实时检测植物体叶片的葡萄糖浓度,为了解霜霉病侵染黄瓜的抗病状态提供支持。【方法】采用双通道丝网印刷电极为基底电极,使用1 mg/mL氯金酸溶液在传感器表面生成金纳米颗粒用于提高其导电性及活性面积,将5 mg/mL壳聚糖和180 mg/mL葡萄糖氧化酶混合以提高酶的固载量和活性,滴加10%的Nafion水溶液以增加对阳离子的选择性并提高电极稳定性。在此基础上研制出一种适用于活体检测植物扁平叶片中葡萄糖浓度的双通道电化学传感器。采用扫描电镜、X射线能谱、红外光谱、循环伏安法及电化学阻抗谱,对所制备传感器进行形貌、结构及电化学的表征;基于优化条件,对所制备的葡萄糖传感器进行性能测试;进一步使用黄瓜果汁进行传感器的加标试验,并应用该传感器对正常与带菌斑黄瓜叶片的葡萄糖浓度进行测定和比较。【结果】所制备植物叶片葡萄糖活体检测传感器的形貌、结构及电化学表征结果显示电极制备成功,其对葡萄糖具有良好的催化效果,反应为扩散控制过程;所设计的植物活体叶片葡萄糖检测双通道传感器的线性检测范围为1～150 mmol/L,检出限为0.784 mmol/L。采用3种不同浓度黄瓜果汁进行加标回收试验,回收率为97.17%~103.05%,相对标准误差≤ 4.52%,表明该传感器准确可靠。应用所制备的传感器检测结果表明,在双通道检测条件下,正常黄瓜叶片和霜霉菌侵染黄瓜叶片在通道1的葡萄糖浓度分别为（8.03±0.96）和（28.71±2.03） mmol/L,在通道2的葡萄糖浓度分别为（12.15±1.46）和（25.57±1.81） mmol/L,霜霉菌侵染黄瓜叶片的葡萄糖浓度较相应正常叶片增长了257.53%（通道1之间）和110.45%（通道2之间）,表明不同生理条件下同种植物叶片的葡萄糖浓度因为霜霉菌侵染而存在显著差异（P＜0.05）;病斑区域葡萄糖浓度的检测结果还显示,通道1的葡萄糖浓度高于通道2,表明通道1检测的侵染区域抗病响应迅速,侵染区相应代谢物质会发生明显变化导致邻近区域的葡萄糖逐步出现积累。【结论】所制备的传感器能够检测植物在不同生理环境下的葡萄糖浓度,可为活体植物叶片葡萄糖的实时检测提供技术支持。     

多效唑对芋种质离体保存影响的初步研究

以荔浦芋无菌试管苗为材料,研究了试管苗保存过程中不同浓度多效唑（PP粥）对试管苗保存效果的影响。研究结果表明,多效唑对芋种质资源的离体保存有显著影响,在光照时间14～16h／d,光照强度1500lx,室温（26＋2）℃的培养条件下,在MS＋6-BA2．5mg／L＋NAA0．02mg／L＋30g/L蔗糖的培养基中添加0．6mg／LPP粥,试管苗可保存240d,存活率为89．7％,且恢复生长后试管苗形态正常、长势良好,与对照株无明显差异。