Ai-Rong Li

Genetic variation and conservation of Changnienia amoena, an endangered orchid endemic to China

Changnienia amoena is a diploid and self-compatible orchid endemic to China. This species is in great danger of extinction with its current distribution being highly fragmented and discontinuous. This study investigated the level and apportionment of genetic diversity of this species using RAPD technique. Based on 119 discernible DNA fragments generated by 16 primers, an intermediate level of genetic diversity was found at the species level with the percentage of polymorphic bands (P) of 76.5%, expected heterozygosity (He) of 0.194. However, the genetic diversity at the population level was significantly lower (P=37.2%, He=0.120) compared with the average of other species with similar life history characteristics. A high level of population differentiation was detected with 43.8% variation resided among populations as measured by AMOVA. It is noteworthy that as much as 49.2% of the total diversity could be attributed to difference among populations when five populations from an area of approximately 60 × 25 km in the Xinning County were considered. These results, in conjunction with other evidence from pollination and ecological studies, suggest that the low within-population but high among-population variation in C. amoena is likely due to the small population sizes and local extinction because of habitat destruction and loss. The restricted gene flow is probably another factor contributing to the genetic structure found in C. amoena. Based on these findings, we proposed conservation managements for this endangered species, including habitat protection along with the protection of their pollinators, artificial pollination as well as ex situ conservation.

Mycorrhizal and dark septate endophytic fungi of Pedicularis species from northwest of Yunnan Province, China

Colonization of mycorrhizal and root endophytic fungi in 14 Pedicularis species from northwest of Yunnan Province, southwest China, was examined. These species included: Pedicularis gracilis Wall., Pedicularis longipes Maxim., Pedicularis axillaris Franch., Pedicularis cephalantha Franch., Pedicularis tenuisecta Franch., Pedicularis tapaoensis Tsoong, Pedicularis likiangensis Franch., Pedicularis dichotoma Bonati, Pedicularis yui Li, Pedicularis rhinanthoides Schrenk, Pedicularis rex C.B. Clarke, Pedicularis longiflora Rudolph., Pedicularis siphonantha Don, and Pedicularis oxycarpa Franch., among which nine are endemic to China (one to Yunnan). Three types of potentially beneficial fungi associated with roots of these species were observed, namely, arbuscular mycorrhizal fungi, ectomycorrhizal fungi, and dark septate endophytic fungi (DSEF), with DSEF as the most common colonizers. An unexpected high colonization level was detected in this hemiparasitic genus. Of the 19 sampling sites examined, 10 gave colonization frequency of above 50% and 6 showed a colonization index of above 50. Heavy colonization suggested a significant ecological role of these fungi and their potential to be applied to successful cultivation of these intractable plants.

Arbuscular mycorrhizal fungi may serve as another nutrient strategy for some hemiparasitic species of Pedicularis (Orobanchaceae)

As an important component of plant kingdom, parasitic plants have intrigued many scientists with their heterotrophic strategy. Numerous investigations have been carried out for a better understanding of interactions between parasitic plants and their hosts. Nevertheless, studies on parasitic plants from a mycorrhizal perspective are lacking, largely because of the notion that parasitic plants do not form mycorrhizal associations. Although long being regarded as nonmycorrhizal, some Pedicularis species are recently found to be heavily colonized by mycorrhizal fungi. Because the precise information about parasitism of Chinese Pedicularis has been lacking, we surveyed both the mycorrhizal status and parasitism of 29 Pedicularis species from the northwest of Yunnan Province, China, to test the hypothesis that some Pedicularis may be mycorrhizal and parasitic simultaneously. The majority of studied species were found to be parasitic as well as mycorrhizal. In some cases, parasitic organs and arbuscular mycorrhizal fungi (AMF) were detected in the same rootlets. The results suggest that some Pedicularis species may have another nutrient strategy (e.g., mycotrophy) besides being parasitic. Also, the findings indicate that host plants as well as AMF should be taken into account in cultivation of Pedicularis species.

Host Dependence and Preference of the Root Hemiparasite, Pedicularis cephalantha Franch. (Orobanchaceae)

The interaction between parasitic plants and their hosts is an important topic in both agriculture and ecology. Little, however, is known about that of the hemiparasite Pedicularis. It is essential to understand the host dependence and preference of Pedicularis for successful cultivation of plants in this genus and for understanding the roles they play in natural communities. We tested the effects of two herbaceous host species (Trifolium repens and Polypogon monspeliensis) on the survival and growth performance of Pedicularis cephalantha. Five P. cephalantha seedlings and two host plants were planted in each pot. In the control, no host plants were planted (treatment 1). Host plants were planted in three combinations: only T. repens (treatment 2) or P. monspeliensis (treatment 3) or a mixture of both (treatment 4). The results showed that P. cephalantha performed better in the presence of host plants, and host plants are more essential to P. cephalantha for proper development than for survival. The grass host P. monspeliensis proved to be a better host plant for P. cephalantha than the legume host T. repens. The high dependence of P. cephalantha on host plants and its host preference were demonstrated in this study. This is the first report of the performance of Chinese Pedicularis species in cultivation throughout all life stages (from seeds to seeds).

Arbuscular mycorrhizal fungi: potential biocontrol agents against the damaging root hemiparasite Pedicularis kansuensis?

Spatial expansion of root hemiparasitic Pedicularis kansuensis in Bayanbulak Grassland of Xinjiang Uygur Autonomous Region (China) has caused great loss of herbage yield and has threatened the local livestock industry. Current management practices using manual eradication and chemical control have been proved problematic. Arbuscular mycorrhizal (AM) fungi have been suggested to be potential biocontrol agents against a number of plant pests, but experimental evidence is lacking against weedy P. kansuensis. In this study, we tested the hypothesis that inoculation with AM fungi will cause growth depression in P. kansuensis and reduce its damage to host plants. Based on the confirmation of AM status and host community of the hemiparasite in the field, a pot cultivation experiment was conducted to test the influence of an AM fungus (Glomus mosseae) on growth of P. kansuensis and the parasitized host (Elymus nutans). AM colonization was observed in roots of P. kansuensis, but the levels were much lower than those of its adjacent host species. A negative correlation between AM levels and the numbers of haustoria was detected for the field samples of the hemiparasite. Strong suppression of haustorium formation, a significant reduction in plant dry weight (DW), as well as marked reduction in the survival rate of P. kansuensis after inoculation with AM fungi was observed. In contrast, inoculation with G. mosseae increased root DW and whole plant DW of parasitized host plants. Our findings demonstrated significantly repressive effects of AM fungi on growth performance of P. kansuensis with and without the presence of a host. The potential of AM fungi as biocontrol agents against the damaging hemiparasite was confirmed.

Two sympatric root hemiparasitic Pedicularis species differ in host dependency and selectivity under phosphorus limitation

Parasitic biology of Pedicularis L. (Orobanchaceae) has been underinvestigated despite its wide distribution and potential ecological significance. To better understand the parasitic aspects of the root hemiparasites, host-parasite interactions were investigated with two sympatric Pedicularis species, Pedicularis rex C. B. Clarke and Pedicularis tricolor Hand.-Mazz., at two developmental stages. Plant DW, shoot phosphorus (P) content, root:shoot ratio and number of haustoria were measured in Pedicularis grown with either a host plant or a plant of its own species in pot experiments. In addition, effects of parasitism and intraspecific competition on growth and biomass allocation in four host species belonging to three major functional groups (grasses, legumes and forbs) were investigated. The two Pedicularis species showed obvious host preference, but preferred different host species. Interactions between Pedicularis and their hosts depended on both species identity and developmental stages of the partners. Overall, P. rex showed much weaker host dependency and less damage to hosts than P. tricolor. Interspecific variations were observed among different host species in their responses to intraspecific competition and parasitism. We concluded that different Pedicularis-host pairs showed different interaction patterns. Sympatric Pedicularis may have differential influence on plant community structure and productivity.

Host shoot clipping depresses the growth of weedy hemiparasitic Pedicularis kansuensis

Root hemiparasitic plants show optimal growth when attached to a suitable host by abstracting water and nutrients. Despite the fact that damage to host plants in the wild occurs frequently in various forms (e.g. grazing), effects of host damage on growth and physiological performance of root hemiparasites remain unclear. In this study, host shoot clipping was conducted to determine the influence of host damage on photosynthetic and growth performance of a weedy root hemiparasite, Pedicularis kansuensis, and its interaction with a host, Elymus nutans. Photosynthetic capacity, tissue mineral nutrient content and plant biomass of P. kansuensis were significantly improved when attached to a host plant. Host clipping had no effect on quantum efficiency (ΦPSII), but significantly reduced the growth rate and biomass of P. kansuensis. In contrast, clipping significantly improved photosynthetic capacity and accumulation of potassium in E. nutans. No significant decrease in biomass was observed in clipped host plants. By changing nutrient absorption and allocation, clipping affected the interaction between P. kansuensis and its host. Our results showed that host clipping significantly suppressed the growth of weedy P. kansuensis, but did not affect biomass accumulation in E. nutans. We propose that grazing (a dominant way of causing host damage in the field) may have a potential in the control against the weedy hemiparasite.

Nutrient requirements differ in two Pedicularis species in the absence of a host plant: implication for driving forces in the evolution of host preference of root hemiparasitic plants

BACKGROUND AND AIMS Facultative root hemiparasitic plants generally have a wide host range, but in most cases show an obvious host preference. The reasons for the marked difference in growth performance of hemiparasites when attached to different hosts are not fully understood. In this study, the hypothesis was tested that hemiparasites showing a preference for different hosts have different nutrient requirements. METHODS Two facultative root hemiparasitic Pedicularis species (P. rex and P. tricolor) with a different host dependency and preference were used to test their responses to inorganic solutes. The effects of nitrogen, phosphorus and potassium on growth of the hemiparasitic plants not attached to a host were determined, using an orthogonal design in pot cultivation under greenhouse conditions. Variables including biomass, shoot nutrient concentration, root:shoot (R:S) ratios and the number of haustoria were measured. KEY RESULTS As in autotrophic plants, nutrient deficiency reduced dry weight (DW) and nutrient concentrations in the root hemiparasites. Nitrogen and phosphorus significantly influenced growth of both Pedicularis species, while potassium availability influenced only shoot DW of P. rex. Nitrogen had far more effect on growth of P. rex than on P. tricolor, while phosphorus deficiency caused more marked growth depression in P. tricolor than in P. rex. Pedicularis rex grew faster than P. tricolor in a range of nutrient supplies. Different patterns of biomass allocation between the two Pedicularis species were observed. While P. rex invested more into roots (particularly fine rootlets) than P. tricolor, the number of haustoria produced by P. rex was relatively much lower than that produced by P. tricolor, which had a much smaller root system. CONCLUSIONS The two Pedicularis species differ in nutrient requirements and biomass allocation. Distinct interspecific traits in growth and nutrient requirements can be driving forces for the differential interactions between hemiparasites and their hosts.

Long-Distance Dispersal after the Last Glacial Maximum (LGM) Led to the Disjunctive Distribution of Pedicularis kansuensis (Orobanchaceae) between the Qinghai-Tibetan Plateau and Tianshan Region

Quaternary climate fluctuations have profoundly affected the current distribution patterns and genetic structures of many plant and animal species in the Qinghai-Tibetan Plateau (QTP) and adjacent mountain ranges, e.g. Tianshan (TSR), Altay, etc. In this greater area disjunct distributions are prominent but have nevertheless received little attention with respect to the historical processes involved. Here, we focus on Pedicularis kansuensis to test whether the current QTP and TSR disjunction is the result of a recent Holocene range expansion involving dispersal across arid land bridge(s) or a Pleistocene range fragmentation involving persistence in refugia. Two chloroplast DNA spacers were sequenced for 319 individuals from 34 populations covering the entire distribution range of this species in China. We found a total of 17 haplotypes of which all occurred in the QTP, and only five in the TSR. Overall genetic diversity was high (HT = 0.882, HS = 0.559) and higher in the QTP than in the TSR. Genetic differentiation among regions and populations was relatively low (GST = 0.366) and little evidence for a phylogeographic pattern emerged. The divergence times for the four main lineages could be dated to the early Pleistocene. Surprisingly, the two ubiquitous haplotypes diverged just before or around the Last Glacial Maximum (LGM) and were found in different phylogenetic lineages. The Species Distribution Model suggested a disappearance of P. kansuensis from the TSR during the LGM in contrast to a relatively constant potential distribution in the QTP. We conclude that P. kansuensis colonized the TSR after the LGM. The improbable long-distance dispersal by wind or water across arid land seed flow may well have had birds or men as vector.

N-P Fertilization Inhibits Growth of Root Hemiparasite Pedicularis kansuensis in Natural Grassland

Fertilization has been shown to affect interactions between root hemiparasitic plants and their host plants, alleviating damage to the hosts by parasitism. However, as a majority of studies were conducted in pot cultivation, the influence of fertilizer application on root hemiparasites and the surrounding plant community in field conditions as well as relevant mechanisms remain unclear. We manipulated soil nutrient resources in a semi-arid subalpine grassland in the Tianshan Mountains, northwestern China, to explore the links between fertilization and plant community composition, productivity, survival, and growth of a weedy root hemiparasite (Pedicularis kansuensis). Nitrogen (at a low rate, LN, 30 kg N ha-1 year-1 as urea; or at a high rate, HN, 90 kg N ha-1 year-1 as urea) and phosphorus [100 kg ha-1 year-1 as Ca(H2PO4)2⋅H2O] were added during two growing seasons. Patterns of foliar nutrient balances were described with isometric log ratios for the different plant functional groups receiving these fertilization regimes. Fertilization with LN, HN, and P reduced above-ground biomass of P. kansuensis, with above-ground biomass in the fertilization treatments, respectively, 12, 1, and 39% of the value found in the unfertilized control. Up to three times more above-ground biomass was produced in graminoids receiving fertilizers, whereas forb above-ground biomass was virtually unchanged by the fertilization regimes and forb species richness was reduced by 52% in the HN treatment. Fertilization altered foliar nutrient balances, and distinct patterns emerged for each plant functional group. Foliar [C | P,N] balance in the plant community was negatively correlated with above-ground biomass (P = 0.03). The inhibited competitiveness of P. kansuensis, which showed a much higher [C | P,N] balance, could be attributed to reduced C assimilation rather than mineral nutrient acquisition, as shown by significant increase in foliar N and P concentrations but little increase in C concentration following fertilization.