Planta

Hand-held Raman spectroscopy can be used for highly accurate differentiation between young male and female hemp plants. This differentiation is based on significantly different concentration of lutein in these plants. Last year, a global market of only industrial hemp attained the value of USD 4.7 billion. It is by far the fastest growing market with projected growth of 22.5% between 2021 and 2026. Hemp (Cannabis sativa L.) is a dioecious species that has separate male and female plants. In hemp farming, female plants are strongly preferred because male plants do not produce sufficient amount of cannabinoids. Male plants are also eliminated to minimize a possibility of uncontrolled cross-fertilization of plants. Silver treatments can induce development of male flowers on genetically female plants in order to produce feminized seed. Resulting cannabinoid hemp production fields should contain 100% female plants. However, any unintended pollination from male plants can produce unwanted males in production fields. Therefore, there is a growing demand for a label-free, non-invasive, and confirmatory approach that can be used to differentiate between male and female plants before flowering. In this study, we examined the extent to which Raman spectroscopy, an emerging optical technique, can be used for the accurate differentiation between young male and female hemp plants. Our findings show that Raman spectroscopy enables differentiation between male and female plants with 90% and 94% accuracy on the level of young and mature plants, respectively. Such analysis is entirely non-invasive and non-destructive to plants and can be performed in seconds using a hand-held spectrometer. High-performance liquid chromatography (HPLC) analysis and collected Raman spectra demonstrate that this spectroscopic differentiation is based on significantly different concentrations of carotenoids in male vs female plants. These findings open up a new avenue for quality control of plants grown in both field and a greenhouse.

Many plants contain ribosome-inactivating proteins (RIPs) which are either single enzymatically active polypeptides (type-1 RIPs) or heterodimers (type-2 RIPs) composed of an A-chain, functionally equivalent to a type-1 RIP, which is disulphide bonded to a sugar-binding B-chain. Much attention has focused on the use of RIPs as components of immunotoxins or, more recently, as antiviral agents. In contrast, relatively little is known about either the synthesis and targeting of RIPs or their role within plants. In this study the cellular and subcellular distributions of saporins, the type-1 RIPs from soapwort, have been determined in seeds using immunogold labelling. Saporins were present in the seed storage tissue (perisperm), but are not synthesised in the developing embryo, demonstrating that the expression of saporin genes is subject to tissue-specific control. Within the perisperm, saporin was found in extracellular spaces, in the paramural region between the primary wall and plasmalemma and within the vacuole. In addition, saporin was localised in leaf intercellular spaces. This dual localisation, both vacuolar and extracellular, is significantly different from the localisation of ricin, a type-2 RIP found in castor beans, which is targeted to endosperm protein bodies, and to pokeweed antiviral protein which accumulates in the cell wall matrix of leaf mesophyll cells.

Hand-held Raman spectroscopy is a potential tool for a confirmatory, non-invasive, and non-destructive detection and identification of rose rosette disease. Using this spectroscopic approach, structural changes in roses that are associated with this viral infection can be revealed. The commercial rose shrub industry in the United States is one of the largest of its kind. All commercial rose varieties are susceptible to rose rosette disease (RRD), a deadly viral disease vectored by eriophyid mites. This disease is typically diagnosed visually and/or by PCR-based detection assays. The present work demonstrates that Raman spectroscopy can detect RRD in intact leaf tissue. It is shown that chemometric analysis can distinguish between spectra collected from symptomatic and asymptomatic tissue, as well as between healthy and asymptomatic tissue. This method will be useful as an initial screen for RRD prior to PCR analysis to help conserve reagents and save time.

Sorbitol metabolism plays multiple roles in many plants, including energy and carbon enrichment, effective defence against various stresses and other emerging specific roles. The underlying mechanisms are, however, incompletely understood. This review provides the current state-of-the-art, highlights missing knowledge and poses several remaining questions. The basic properties of sugar alcohols are summarised and pathways of sorbitol metabolism, including biosynthesis, degradation and key enzymes are described. Sorbitol transport within the plant body is discussed and individual roles of sorbitol in different organs, specific cells or even cellular compartments, are elaborated, clarifying the critical importance of sorbitol allocation and distribution. In addition to plants that accumulate and transport significant quantities of sorbitol (usual producers), there are some that synthesize small amounts of sorbitol or only possess sorbitol metabolising enzymes (non-usual producers). Modern analytical methods have recently enabled large amounts of data to be acquired on this topic, although numerous uncertainties and questions remain. For a long time, it has been clear that enriching carbohydrate metabolism with a sorbitol branch improves plant fitness under stress. Nevertheless, this is probably valid only when appropriate growth and defence trade-offs are ensured. Information on the ectopic expression of sorbitol metabolism genes has contributed substantially to our understanding of the sorbitol roles and raises new questions regarding sorbitol signalling potential. We finally examine strategies in plants producing sorbitol compared with those producing mannitol. Providing an in-depth understanding of sugar alcohol metabolism is essential for the progress in plant physiology as well as in targeted, knowledge-based crop breeding.

Chemical gradients and structural features within the pistil have been previously proposed as factors determining the directionality of pollen tube growth. In this study, we examine the behavior of pollen of eight species germinated in a dynamic oxygen gradient. While the germination rates of some species decreased directly with decreasing oxygen tension, other species showed no decrease in germination at oxygen tensions as low as 2 kPa. In one species, germination was consistently greater at decreased oxygen tensions than at ambient atmospheric levels. In three of the eight species tested, the developing pollen tube showed clear directional growth away from the more-oxygenated regions of the growth medium, while in one species growth was towards the more-oxygenated region. The remaining four species showed random tube growth. The pattern of oxytropic responses among the taxa suggests that this tropic behavior is both widespread and phylogenetically unpredictable.

Microspores of Brassica napus L. cv. Topas, undergo embryogenesis when cultured at 32.5 °C for the first 18–24 h and then at 25 °C. The first division in heat-treated microspores is a symmetric division in contrast to the asymmetric division found after the first pollen mitosis in-planta or in microspores cultured continuously at 25 °C. This asymmetric division is unique in higher plants as it results in daughter cells separated by a non-consolidated wall. The cytoskeleton has an important role in such morphological changes. We examined microtubule (MT) organization during the first 24 h of heat induction in the embryogenic B. napus cv. Topas and the non-embryogenic B. napus breeding line 0025. Preprophase bands (PPBs) of MTs appeared in cv. Topas microspores in late uninucleate microspores and in prophase figures after 4–8 h of heat treatment. However, more than 60% of the PPBs were not continuous bands. In contrast, PPBs were never observed in pollen mitosis; MT strands radiated from the surface of the nuclear envelope throughout microspore maturation to the end of prophase of pollen mitosis I, during in-planta development and in microspores cultured at 25 °C. Following 24 h of heat treatment, over 95% of the microspores appeared to have divided symmetrically as indicated by the similar size of the daughter nuclei, but only 7–16% of the microspores eventually formed embryos. Discontinuous walls were observed in more than 50% of the divisions and it is probable that the discontinuous PPBs gave rise to such wall abnormalities which may then obstruct embryo development. Preprophase bands were not formed in heat-treated microspores of the non-embryogenic line 0025 and the ensuing divisions showed discontinuous walls. It is concluded that the appearance of PPBs in heat-induced microspores marks sporophytic development and that continuous PPBs are required for cell wall consolidation and embryogenesis. It follows that induced structures with two equally condensed nuclei, do not necessarily denote symmetric divisions.