Simple sequence repeat (SSR) markers are among more reliable markers widely used in population genetics and traditionally need intensive labor to develop. High-throughput sequencing (HTS) has enabled researches in screening whole DNA in order to develop dozens of reliable SSR markers to pave route for future studies. Annual medics ( Medicago spp.) are specifically beneficial in agriculture, genetics, and pastureland management plans. In order to facilitate study of annual medics using SSRs and to test their effectiveness, HTS screening of whole DNA resulted in developing of hundreds of primer pairs published in this paper. Subsequent testing of candidate primers on Iranian M. orbicularis germplasm was narrowed down to six more variable primer pairs that generated 28 polymorphic bands with the allele size of 200–310 bp. Eleven populations were examined using our SSR markers. The average polymorphic information content was very high (> 0.90) for the primer combinations. Genetic diversity parameters were measured for all populations. A high level of polymorphism of 75% within populations was observed. We hypothesized that the self-pollinating reproductive system of M. orbicularis has led to the reduction of gene flow between populations and ultimately the low diversity among populations (24%). Different genotypes observed by structure analysis as well as regression and correlation analyses showed that genetical similarity was congruent with the geographical proximity. Our results suggest that our HTS-generated SSR markers can be used for the further study of other annual medics.

ABSTRACT Species of invertebrate animals, notably insects, are undergoing an alarmingly high rate of extinction, coupled with minimal support for their protection, even from the world’s leading conservation organisations. This is intolerable, as invertebrates constitute over 95% of the world’s species, have indispensable economic values and provide ecological services without which life on earth would virtually cease. Much of the lack of public and governmental support for invertebrate conservation is due to the abhorrent tiny pests that have persuaded most people that ‘bugs’ are bad and consequently the only species worthy of support are the charismatic superstar mammals like pandas and tigers that currently are the mainstays of biodiversity fundraising. Just as these respected, highly attractive icons are effective ambassadors of biodiversity conservation, so certain detested pests have poisoned the public image of invertebrates, and indeed have made it seem to many that most wildlife is hostile. The ‘dirty dozen’ bugs that particularly are a hindrance to improving public investment in biodiversity are: bedbugs, clothes moths, cockroaches, fleas, houseflies, leeches, lice, locusts, mosquitoes, spiders, termites and ticks. Except for spiders, these species, admittedly, are responsible for enormous damage to health and economic welfare. Nevertheless, this paper shows that most have at least some compensating values, their harm has often been exaggerated and all have related species that are good citizens. Six of the dozen ‘least wanted’ invertebrates highlighted are blood parasites of humans, and these ‘bad apples’ are very hard to defend since parasitism seems abhorrent. Remarkably, however, at least half of the world’s tens of millions of species are also parasites, and without them most ecosystems would be in danger of collapse. To improve invertebrate conservation, it is advisable that efforts be made to educate the public regarding their importance. Since prejudices against ‘bugs’ are primarily acquired during childhood, special attention is needed to persuade the young that most invertebrates are harmless, valuable and entertaining. Recent advances in genetic engineering (‘synthetic biology’, ‘genetic drives’) have led to very serious consideration of deliberately eliminating the world’s worst pests of humans. While these extermination technologies could greatly increase support for invertebrate conservation by annihilating their most despised representatives, the dangers of unforeseen damage to ecosystems and hence to biodiversity are substantial.

In this paper, pod morphological traits of eleven populations of Medicago orbicularis (L.) Bartal., collected from the West, Northwest, and Southwest of Iran, have been assessed. In addition, nuclear ribosomal Internal Transcribed Spacer (nrITS, ITS1+5.8s+ITS2) variation in four populations of M. orbicularis was analyzed. Molecular phylogenetic analysis showed that all accessions of M. orbicularis formed a well-supported monophyletic clade while no geographical patterning was observed. The result of Principle Components Analysis (PCA) and cluster analysis (Ward’s method) (despite a negative Mantel test), suggested a relationship between the morphology and geographical distribution of the populations.Moreover, three distinctive geographical groups were determined using PCA and cluster analysis. Correlation analyses between ecological variables and morphological traits were often negative. However, altitude, average temperature, and average monthly soil temperature showed positive correlations. Wind's relative effect on all morphological traits has also been observed. Two morphological traits- Seeds Total Number (STN) per pod and Fruit Diameter (FDM)-indicated a significant variation. A direct relationship was observed between the two mentioned characters in all populations. Paveh (PVH) and Marivan (MRV) with the highest STN and FDM demonstrated useful potential for breeding and conservation programs in the future. Overall, it could be assumed that the high STN in M. orbicularis (comparison with other species of Medicago studied in Iran, such as M. minima, M. sinskiae, and M. polymorpha), as well as the effect of winds on seeds distribution, are two main influential factors in creating geographical pattern and morphological diversity.

Medicago sinskiae es considerada una especie rara en la flora iraní con únicamente unas pocas citas en las tres últimas décadas. Se han muestreado ocho poblaciones (62 individuos) de M. sinskiae, una población de M. rigidula (siete individuos) y una población de M. constricta (cinco individuos) en el oeste y el suroeste de Irán que han sido analizadas con marcadores microsatélite. Se han utilizado nuevos primers obtenidos con tecnología NGS. Los análisis de PCoA, Clustering y Structure no muestran un patrón geográfico para los clústeres genéticos. Los resultados muestran que M. sinskiae es principalmente una especie autógama. Se asume que los altos niveles de flujo genético (Nm) y la homogeneidad genética poblacional están afectados por una rápida dispersión y un flujo genético no localizado. Recolecciones extensivas realizadas recientemente en el oeste y el suroeste de Iran muestran que el rango de distribución esta especie se está incrementando. Finalmente, nuestros resultados indican que M. sinskiae está diferenciada de las especies M. rigidula y M. constricta en Irán.

Medicago minima (L.) Bartal. is an important rangeland forage and weed, native to Eurasia and Africa but widely distributed elsewhere. Like dozens of its nearest annual Medicago species, it is known to be highly uniform due to extreme inbreeding, and no valid infra-specific taxa meriting recognition have yet been proposed. In this study, 67 individuals from 13 natural populations of M. minima were collected and subjected to genetic diversity evaluation using 6 simple sequence repeat (SSR). Of the genetic variation parameters, expected heterozygosity ( H e ) showed a significant variation from 0.0 in the Meshkinshahr population to 0.337 in the Andimeshk population. The analysis of molecular variance (AMOVA) revealed considerable partitioning of genetic diversity among populations (58%), while intrapopulation diversity accounted for 42%. Moderate to high population genetic differentiation was observed among particular populations, ranging from 0.060 between Marivan and Paveh to 0.562 between Meshkinshahr and Kohdasht populations. A weak total gene flow (0.115) was accompanied by a relatively high PhiPT value (0.617). Structure analysis, principal coordinate analysis, and neighbour-joining analysis, all divided the 13 populations into two genetic groups with significant genetic homogeneity. This discovery of two molecularly distinctive groups of M. minima in Iran, with a limited hybrid zone between them, is remarkable and suggests early divergent speciation.

It appears that just as we humans express both our love for, and hatred against, certain groups within our species, our closest animal relatives, the mammals, also receive very selective admiration and detestation. Mammals such as those included in the ‘charismatic megafauna’ (big and attractive as shown in Figure 1) and the cute & cuddly species (Figure 2) make ideal aids for enlisting conservation support. In parallel, as noted in this paper, certain species of mammals are passionately hated and are often the subjects of extermination campaigns. These hated species are major concerns, and their intrusions into the artificial habitats of people provoke negative sentiments against the world of wildlife from which they originate. We humans wage war against our enemies and our impulse is to remove the objectionable species not just from our urbanized world, but everywhere. However, the most offensive nuisance mammals are superlative competitors and survivors and attempts to eliminate them require such extensive measures that, inevitably, many other species are endangered. What is needed is to find ways of living with these pests that minimize their harmful effects and a key first step is to learn to understand and respect their needs. Towards this goal, this review presents both the negative and positive aspects of our most detested mammalian pests emphasizing that, on balance, their benefits to humans exceeds their harm. Small (2019) explored how the most reviled species of invertebrate animals significantly but irrationally prejudice the public against wildlife and mitigate efforts to rehabilitate and conserve biodiversity. Part 1 of this paper (Small 2020) similarly carried out the same exercise for the most reviled species of ‘lower vertebrates’ (fish, frogs & toads, snakes, and birds). This follow-up review conducts a similar analysis of the most reviled mammals, which we humans arrogantly consider to be the ‘highest’ group of animals because we are the predominant member. Burgin et al. (2018) list 6400 species of living mammals which represent less than 10% of recognized vertebrate species and less than 0.5% of all animal species. Nevertheless, mammals overwhelmingly dominate conservation initiatives. By a considerable margin, the public supports conservation and rehabilitation of certain ‘charismatic’ mammals much more than any other species (Table 1). As with the previous examination of non-mammal vertebrates (Small 2019), the goal is to generate understanding of the economic values and useful roles of the world’s most disliked mammals in order to minimize the disrespect for biodiversity that they generate. The most despised mammals include bats, hyenas, mice, rats, and skunks (Figure 3). Notably, most of these disreputable mammals are no larger than a housecat whereas the most respected mammals are usually huge. Size is one of the characteristics that strongly determines whether a species is liked or disliked by humans (Small 2011, 2012) and even though most mammals are relatively small they are at least as vital to the welfare of the world as the giants. In relation to their relatively small number of species, mammals nevertheless play disproportionately large economic roles in human existence. Domesticated mammals, and in some regions wild mammals, provide food and hides. Livestock mammals furnish most of the world’s meat, milk, leather, and wool (as noted by Thornton 2019, there are currently 1.5 billion cows, 1 billion sheep, and 1 billion pigs in the world). Some species are important beasts of burden for riding, hauling, and ploughing. The dung and urine of livestock provide an agricultural fertilizer that is superior to today’s synthetic versions. Dogs and cats have become the world’s major pets, and dogs are also invaluable working assistants. (Unfortunately most of the approximately 1 billion dogs [Atitwa 2018] and about 600,000 cats [Migiro 2018] in the world are ‘free-range’ or feral,

A comprehensive study was undertaken to understand the genetic status and help to clarify the division of Medicago rigidula (largely European) and Medicago rigiduloides (largely Asian). Genetic diversity parameters collectively suggested a low genetic diversity (avg. Ho, 0.073; He, 0.374) accompanied by high population differentiation (avg. F, 0.832; Gst, 0.362). Structure analysis divided 71 individuals (14 Iranian populations) into two highly distinct genetic groups (K=2) with significant genetic homogeneity. It also indicated the strong effect of the selfing mating-system as the main reason for the genetic diversity status and population structure. The population grouping was strongly confirmed by various clustering methods. Populations from north and northwestern Iran made up a distinctive genetic group corresponding to M. rigidula while the second group corresponding to M. rigiduloides harboured the western and two of the northwestern populations. The outcomes of this study provide the first reliable molecular evidence supporting the M. rigidula-M. rigiduloides separation previously suggested by morphology.

Burr medic (Medicago polymorpha L.), a native of the Mediterranean Basin, has become a worldwide weed. Pasture cultivars have been selected, especially in Australia. The species is extremely variable, and there is little agreement on its infraspecific classification. Towards a better understanding of its variation pattern, present study examined population genetic structure and genetic diversity in Iran, employing co-dominant SSR markers. Seventy-one randomly collected plants from 14 geographical populations were sampled. Six primer pairs generated 21 polymorphic bands. The average polymorphic information content was high (0.923). Genetic diversity parameters were determined in these populations. STRUCTURE analysis and K-Means clustering identified two and three gene pools in the country. Mantel test results revealed isolation by distance among the studied populations. The cluster analysis showed correlation between genetic and geographical distance. AMOVA revealed a significant genetic difference among populations and showed that 46% of total genetic variation was due to among population diversity. These data may be useful for future breeding and conservation of this important and potentially agricultural plant.

The genus Empetrum (Ericaceae) is controversially classified taxonomically. It is conservatively treated as comprising one variable widespread circumboreal/circumarctic species, E. nigrum, usually known as black crowberry (although there are other fruit colors), and a comparatively localized circumantarctic species, E. rubrum, called red crowberry. For millennia in the Northern Hemisphere crowberries have been a valuable source of berries for Indigenous Peoples, and indeed Empetrum is one of the most important berry crops of the Arctic. It has recently begun to be marketed as a commercial processed fruit crop, with increasing evidence of possessing phenolic compounds of high value for nutrition and medicine. Ecologically, Empetrum is a keystone species, sustaining numerous birds and mammals, and dominating many tundra and heathland ecosystems through allelopathic toxins that exclude competitive plants. With climate change expected to greatly alter the northern world in the near future, there is considerable concern about the welfare of Empetrum.

Biodiversity is vital to the welfare and survival of humans, but public support for conservation of most animal species is appallingly limited. Vertebrates make up less than 5% of the world’s documented animal species, but are viewed far more sympathetically than invertebrates. This is because humans are empathetic with the appearance and behaviour of many of them, particularly the charismatic superstars like pandas and tigers that currently are the mainstays of biodiversity fundraising. Conversely, just as such attractive icons are effective ambassadors of biodiversity conservation, so certain detested and sometimes dangerous vertebrate pests have greatly compromised the public image of biodiversity. Some of these species, admittedly, are responsible for significant damage to health and economic welfare. Nevertheless, this paper shows that all play important ecological roles, they have compensating economic values, their harm has often been exaggerated, and their very negative public images are undeserved. This first installment deals with the most reviled ‘lower’ vertebrate species: sharks (representing fish); frogs and toads (representing amphibians); snakes (representing reptiles); and vultures (representing birds). The next contribution will deal with mammals.

Cannabis sativa has been domesticated for stem fibre and oilseed (the two classes are both low in the euphoric cannabinoid THC and called “hemp”), and marijuana (high in THC), and also occurs as weedy, ruderal plants. Achenes (“seeds”) from herbarium collections representative of these classes were assessed for morphological characters and pericarp resistance to fracture. In contrast to ruderal plants, domesticated plants (both hemp and marijuana) possessed achenes that were significantly longer, heavier, covered with a less adherent perianth, and lacking a pronounced basal attenuation. All of these characteristics reflect traits that are advantageous in domesticated plants and are consistent with the “domestication syndrome” found in propagules of other crops. Marijuana achenes, in comparison with hemp achenes, tended to be about 26% shorter and about 32 shades darker (on a 256-bit grayscale). Achenes of fibre cultivars proved to be about 19% longer than the achenes of oilseed cultivars. Achenes of dioecious oilseed cultivars proved to be about 6% longer than the achenes of monoecious oilseed cultivars. The pericarps of hemp seeds were about 26% and about 15% more resistant to fracture than those of ruderal and marijuana plants, respectively.

Two kinds of drug-type Cannabis gained layman’s terms in the 1980s. “Sativa” had origins in South Asia (India), with early historical dissemination to Southeast Asia, Africa, and the Americas. “Indica” had origins in Central Asia (Afghanistan, Pakistan, Turkestan). We have assigned unambiguous taxonomic names to these varieties, after examining morphological characters in 1100 herbarium specimens, and analyzing phytochemical and genetic data from the literature in a meta-analysis. “Sativa” and “Indica” are recognized as C. sativa subsp. indica var. indica and C. sativa subsp. indica var. afghanica, respectively. Their wild-growing relatives are C. sativa subsp. indica var. himalayensis (in South Asia), and C. sativa subsp. indica var. asperrima (in Central Asia). Natural selection initiated divergence, driven by climatic conditions in South and Central Asia. Subsequent domestication drove further phytochemical divergence. South and Central Asian domesticates can be distinguished by tetrahydrocannabinol and cannabidiol content (THC/CBD ratios, ≥7 or <7, respectively), terpenoid profiles (absence or presence of sesquiterpene alcohols), and a suite of morphological characters. The two domesticates have undergone widespread introgressive hybridization in the past 50 years. This has obliterated differences between hybridized “Sativa” and “Indica” currently available. “Strains” alleged to represent “Sativa” and “Indica” are usually based on THC/CBD ratios of plants with undocumented hybrid backgrounds (with so-called “Indicas” often delimited simply on possession of more CBD than “Sativas”). The classification presented here circumscribes and names four taxa of Cannabis that represent critically endangered reservoirs of germplasm from which modern cannabinoid strains originated, and which are in urgent need of conservation.

Unsustainable agriculture, particularly with respect to crops, is the chief threat to planetary resources, including the atmosphere, water, soil, landscapes and biodiversity (DeWitt 2009). For several decades, the cannabis plant (Cannabis sativa) has been unrealistically extolled as the potential saviour of habitats, ecosystems and the environment, because of its claimed ecological benefits (Herer 1998). As noted below, this is highly misleading: as fibre and oilseed crops, the plant indeed has admirable environmentally-friendly properties, but as a source of medicinal and recreational drugs, its cultivation is significantly consumptive and polluting. Currently, huge expenditures of energy are associated with growing marijuana plants in high-technology greenhouses (Figure 1) rather than as nature intended: outdoors. While the issue of greenhouse production of marijuana has recently become prominent because of enormous expansion of the legitimate marijuana industries, the ecological justification of greenhouse cultivation in general is very important for the welfare of the planet and its living inhabitants. As discussed in this review, greenhouses can be environmentally benign or unacceptably harmful. Marijuana represents the most rapidly expanding industry in the Western world (Borchardt 2017), with the potential to generate trillions of dollars of business annually in the near future. Immense fortunes are now being invested in the cultivation of marijuana plants, and much of this is based on growing them in energyconsuming greenhouses which can contribute significantly to atmospheric deterioration and indirectly to endangerment of biodiversity. This contribution examines the ways that cultivation of marijuana plants may be ecologically damaging, with particular emphasis on the role of greenhouses. As detailed elsewhere (Montford and Small 1999a, 1999b; Small and Catling 2009; Small 2015), when grown for fibre or for oilseed, C. sativa is one of the world’s most ecologically beneficial crops, and it is perfectly feasible to grown the plant for production of marijuana in an equally sustainable fashion.

Ivies (species of Hedera) were universally admired ornamental vines until several decades ago, and to this day most horticulturalists continue to defend their use. However, Ivies have escaped in wi...

After a century of banishment, both euphoric (“marijuana”) and non-euphoric (“industrial hemp”) classes of Cannabis sativa are attracting billions of dollars of investment as new legitimate crops. Most domesticated C. sativa is very tall, a phenotype that is desirable only for hemp fibre obtained from the stems. However, because the principal demands today are for chemicals from the inflorescence and oilseeds from the infructescence, an architecture maximizing reproductive tissues while minimizing stems is appropriate. Such a design was the basis of the greatest short-term increases in crop productivity in the history of agriculture: the creation of short-stature (“semi-dwarf”), high-harvest-index grain cultivars, especially by ideotype breeding, as demonstrated during the “Green Revolution.” This paradigm has considerable promise for C. sativa. The most critical dwarfing character for breeding such productivity into C. sativa is contraction of internodes. This reduces stem tissues (essentially a waste product except for fibre hemp) and results in compact inflorescences (which, on an area basis, maximize cannabinoid chemicals) and infructescences (which maximize oilseed production), as well as contributing to ease of harvesting and efficiency of production on an area basis. Four sources of germplasm useful for breeding semi-dwarf biotypes deserve special attention: (1) Naturally short northern Eurasian wild plants (often photoperiodically day-neutral, unlike like most biotypes) adapted to the stress of very short seasons by maximizing relative development of reproductive tissues. (2) Short, high-harvest-index, oilseed plants selected in northern regions of Eurasia. (3) “Indica type” marijuana, an ancient semi-dwarf cultigen tracing to the Afghanistan-Pakistan area. (4) Semi-dwarf strains of marijuana bred illegally in recent decades to avoid detection when grown clandestinely indoors for the black market. Although the high THC content in marijuana strains limits their usage as germplasm for low-THC cultivars, modern breeding techniques can control this variable. The current elimination of all marijuana germplasm from breeding of hemp cultivars is short-sighted because marijuana biotypes possess a particularly wide range of genes. There is an urgent need to develop public gene bank collections of Cannabis.

The principal economic species of the genus Melilotus are white sweet-clover (Melilotus albus) and the extremely similar yellow sweet-clover (M. officinalis). Although they are widely recognized as distinct species, some influential references in North America reduce the former to a subspecific rank or even merely a conspecific synonym of the latter. Given their importance and the large numbers of germplasm collections, the doubt needs to be resolved. This review of relevant published evidence finds that in addition to the difference in floral colour, the traditional segregation of the two as distinct species is best supported by very strong reproductive barriers as well as divergent DNA sequences in three barcoding genes. Additional but weaker confirmation of separateness is provided by studies reporting differences in external morphology, biochemistry, seed protein profiles, karyotype and DNA microsatellites.

Most of the world has become acquainted with the concerns over overfishing and pollution as major threats to the commercial harvest of marine animals and the health of the seas and oceans. This rev...

We report here that small but numerous foliar glands are present near the base of the abaxial leaf blades of hop (Humulus lupulus L.) at about 2.6 times the density found in the more distal regions, and this concentration of glands appear to protect the basal region from damage by insects. Distinctive wild taxa of H. lupulus are native to distinctive regions of the north temperate world. The much higher presence of protective glands in the three native North American Humulus taxonomic varieties in comparison with the varieties native to Europe and Asia suggests specialized leaf feeders in North America have been responsible for the evolution of greater gland density. Commercial domesticated hop cultivars trace most of their germplasm to the European hop (H. lupulus var. lupulus), which possesses a much lower concentration of leaf glands than the native hops of all other regions of the world, and therefore is likely relatively susceptible to damage. We suggest that breeding for increased density of protective foliage glands may be a non-chemical way of alleviating foliar feeding damage in cultivated hops.