Composition of nutrient media, flower bud size, sucrose concentration, heat shock stress, and ethylene inhibitor could have marked effects on microspore embryogenesis. No microspore-derived embryos (MDE) were formed when the microspores were isolated from radish (Raphanus sativus L.) flower buds of 1.0-2.5 mm in size, whereas MDE were formed with microspores isolated from 2.5-4.5 and 4.5-6.5 mm flower buds. The microspores isolated from 2.5-4.5 mm flower buds showed high embryo yields. MDE formation was highest when 150 g L -1 sucrose was added to the half strength Nitsch & Nitsch (NLN) liquid medium, but at sucrose concentrations less than 100 g L -1 there was no MDE formation. Microspores cultured on half strength NLN liquid medium containing 0.05 mg L -1 silver nitrate (AgNO3) produced the most MDE, showing more than two-fold increase in yield compared to those cultured on medium without AgNO3. A heat shock pretreatment of microspores at 32ºC for 24 h gave high-frequency production of MDE when compare to higher or lower temperatures; no MDE were formed at 42.5ºC. The highest yield of MDE was observed when microspores were derived from 2.5-4.5 mm flower buds cultured on half strength NLN medium containing 150 g L -1 sucrose, 0.05 mg L -1 AgNO3, and precultured with heat shock pretreatment of microspores at 32ºC for 24 h, followed by incubation 25ºC for 30 days. A polyploidy test indicated that 19.7% of the microspore-derived plants were doubled haploid, other plants were haploid, and chimeras were haploid and diploid.Additional key words: double haploid, embryogenesis, flower bud, heat shock, Nitsch & Nitsch (NLN) medium, polyploidy testHort. Environ. Biotechnol. 52(5):530-535. 2011.

This study aimed to establish a rapid in vitro plant regeneration method from rhizome buds of Kaempferia parviflora to obtain the valuable secondary metabolites with antioxidant and enzyme inhibition properties. The disinfection effect of silver oxide nanoparticles (AgO NPs) on rhizome and effects of plant growth regulators on shoot multiplication and subsequent rooting were investigated. Surface sterilization of rhizome buds with sodium hypochlorite was insufficient to control contamination. However, immersing rhizome buds in 100 mg L−1 AgO NPs for 60 min eliminated contamination without affecting the survival of explants. The number of shoots (12.2) produced per rhizome bud was higher in Murashige and Skoog (MS) medium containing 8 µM of 6-Benzyladenine (6-BA) and 0.5 µM of Thidiazuron (TDZ) than other treatments. The highest number of roots (24), with a mean root length of 7.8 cm and the maximum shoot length (9.8 cm), were obtained on medium MS with 2 µM of Indole-3-butyric acid (IBA). A survival rate of 98% was attained when plantlets of K. parviflora were acclimatized in a growth room. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) was used to determine the chemical profile of K. parviflora leaf extracts. Results showed that several biologically active flavonoids reported in rhizomes were also present in leaf tissues of both in vitro cultured and ex vitro (greenhouse-grown) plantlets of K. parviflora. We found 40 and 36 compounds in in vitro cultured and ex vitro grown leaf samples, respectively. Greenhouse leaves exhibited more potent antioxidant activities than leaves from in vitro cultures. A higher acetylcholinesterase inhibitory ability was obtained for greenhouse leaves (1.07 mg/mL). However, leaves from in vitro cultures exhibited stronger butyrylcholinesterase inhibitory abilities. These results suggest that leaves of K. parviflora, as major byproducts of black ginger cultivation, could be used as valuable alternative sources for extracting bioactive compounds.

The radish is a delicious, healthy vegetable and an important ingredient to many side dishes and main recipes. However, climate change, pollinator decline, and especially Fusarium wilt cause a significant reduction in the cultivation area and the quality of the radish yield. Previous studies on plant disease identification have relied heavily on extracting features manually from images, which is time-consuming and inefficient. In addition to Red-Green-Blue (RGB) images, the development of near-infrared (NIR) sensors has enabled a more effective way to monitor the diseases and evaluate plant health based on multispectral imagery. Thus, this study compares two distinct approaches in detecting radish wilt using RGB images and NIR images taken by unmanned aerial vehicles (UAV). The main research contributions include (1) a high-resolution RGB and NIR radish field dataset captured by drone from low to high altitudes, which can serve several research purposes; (2) implementation of a superpixel segmentation method to segment captured radish field images into separated segments; (3) a customized deep learning-based radish identification framework for the extracted segmented images, which achieved remarkable performance in terms of accuracy and robustness with the highest accuracy of 96%; (4) the proposal for a disease severity analysis that can detect different stages of the wilt disease; (5) showing that the approach based on NIR images is more straightforward and effective in detecting wilt disease than the learning approach based on the RGB dataset.

RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in eukaryotes, including higher plants. To counteract this, several plant viruses express silencing suppressors that inhibit RNA silencing in host plants. Here, we show that both 2b protein from peanut stunt virus (PSV) and a hairpin construct (designated hp-RDR6) that silences endogenous RNA-dependent RNA polymerase 6 (RDR6) strongly suppress RNA silencing. The Agrobacterium infiltration system was used to demonstrate that both PSV 2b and hp-RDR6 suppressed local RNA silencing as strongly as helper component (HC-Pro) from potato virus Y (PVY) and P19 from tomato bush stunt virus (TBSV). The 2b protein from PSV eliminated the small-interfering RNAs (siRNAs) associated with RNA silencing and prevented systemic silencing, similar to 2b protein from cucumber mosaic virus (CMV). On the other hand, hp-RDR6 suppressed RNA silencing by inhibiting the generation of secondary siRNAs. The small coat protein (SCP) of squash mosaic virus (SqMV) also displayed weak suppression activity of RNA silencing. Agrobacterium-mediated gene transfer was used to investigate whether viral silencing suppressors or hp-RDR6 enhanced accumulations of green fluorescence protein (GFP) and β-glucuronidase (GUS) as markers of expression in leaf tissues of Nicotina benthamiana. Expression of both GFP and GUS was significantly enhanced in the presence of PSV 2b or CMV 2b, compared to no suppression or the weak SqMV SCP suppressor. Co-expression with hp-RDR6 also significantly increased the expression of GFP and GUS to levels similar to those induced by PVY HC-Pro and TBSV P19.