Transcriptome approach to understand the potential mechanisms of resistant and susceptible alfalfa(Medicago sativa L.) cultivars in response to aphid feeding

Plant breeding for resistance to agricultural pests is an essential element in the development of integrated crop management systems, however, the molecular and genetic mechanisms underlying resistance are poorly understood. In this pilot study, we conducted a transcriptomic analysis of a resistant （R） and susceptible （S） variety of alfalfa, with （＋A） or without （-A） aphids （totally four treatments）. We used the resistant cultivar Zhongmu t and the susceptible cultivar Soca. A total of 3 549 mRNAs were differentially expressed, of which 1 738 up-regulated and 1 307 down-regulated genes were identified in S＋A/S-A plants, while 543 up-regulated and 331 down-regulated genes were identified in the R＋A/R-A plants. KEGG analysis mapped 112 and 546 differentially expressed genes to 8 and 17 substantially enriched pathways for Zhongmu 1 and Soca, respectively. Six shared pathways were linked to plant resistance traits, including phenylpropanoid biosynthesis associated with salicylic acid synthesis, and linoleic acid metabolism associated with both jasmonic acid and flavonoid biosynthesis. Ultimately, we proposed a preliminary regulatory mechanism of alfalfa cultivar resistance response to aphids feeding based on transcriptome analyses and published documents.

Including predator presence in a refined model for assessing resistance of alfalfa cultivar to aphids

The aphid quantity ratio（AQR） is defined as the number of aphids on each cultivar divided by the number of aphids on all cultivars. AQR is based on the correlation between aphid populations and their host plants and is an important tool that has been utilized in evaluating Medicago sativa（alfalfa） cultivar resistance to aphids. However, assessment of alfalfa resistance to aphids can be confused by the presence of aphid predators, causing the assessment of plant resistance to aphids to be based on incorrect aphid population data. To refine the AQR and account for the effect of predators on aphid population assessments, we introduced a parameter ‘α＇, corresponding to the predator quantity ratio, and used αAQR as the ratio to quantify aphid populations. Populations of both aphids（4 species） and their predators（12 species） occurring in 28 M. sativa cultivars were sampled over two years at a research station near Cangzhou, Hebei Province, China. Results showed that the most suitable evaluation period was from May to June, as the aphid population was stable during this period. Compared with the AQR method, the predator population numbers based on the αAQR had a significant inverse relationship with aphid population numbers and the 28 cultivars were clustered into three classes： the resistant class, tolerant class, and susceptible class. In addition, 17 cultivars were reassigned when evaluated using αAQR. All numerical values calculated by αAQR were displayed as a Gaussian distribution, which showed that the 28 cultivars could be clustered into nine groups using a median value（±SE） of 1±0.1. Hence, ongoing alfalfa breeding trials will be assessed using the αAQR to establish a robust system that includes agronomic performance parameters in order to generalize the new method for further studies.

Improving a method for evaluating alfalfa cultivar resistance to thrips

The thrips quantity ratio（TQR） model is an important tool for evaluating crop resistance to thrips based on the correlation between thrips quantities and cultivars. Unfortunately, it is inaccurate, and the results appear significantly inconsistent when analysing the same cultivars in the same field study. To improve this model, we first studied the resistance of 28 alfalfa cultivars to thrips in Cangzhou, Hebei Province, north China. The results showed that the most suitable evaluation period was from May to June, as the thrips population was stable during this period. Second, we found that the natural enemy population was significantly positively correlated with the thrips population density（R=0.7275, P〈0.0001）, which might influence resistance estimation. Hence, we introduced a parameter ‘α＇, corresponding to the natural enemy quantity ratio, to eliminate the effect of the natural enemy using ＂αTQR＂. Using the improved method, 28 cultivars were clustered into three classes： the resistant class, sensitive class, and median class. All numerical values were calculated for αTQR displayed as a Gaussian distribution. This information showed that all data should be divided into nine groups using a median value of 1±0.1 with an equal difference of 0.1. Based on the new standard cultivars, Gongnong 1, Alfaking, Cangzhou and Algonquin were classified as highly resistant cultivars; Zhongmu 3, Gongnong 2, Zhongmu 1 and Zhongmu 2 were classified in the resistant group; Queen was classified in the moderately resistant group; Derby, WL354HQ, KRIMA, Apex, 53 HR, SARDI 5 and Farmers Treasure were classified in the median class; WL319HQ, WL343HQ and Sitel were classified as the low sensitive group; WL440 HQ and SARDI7 as the moderately sensitive group; WL168HQ and Sanditi as the sensitive group; and SARDI 10, WL363HQ, FD4, WL323 and SOCA as the highly sensitive group.

Resistance and susceptibility of alfalfa （Medicago sativa L.） cultivars to the aphid Therioaphis maculata （Homoptera： Aphididae）： insect biology and cultivar evaluation

Biology of the aphid Therioaphis maculata was studied on alfalfa （Medicago sativa L.）, including four resistant （Mesa-Sirsa, CUF101, Baker and Lahontan） and two susceptible （ARC and Caliverde） alfalfa cultivars, and one of the most cropped Brazilian cultivars, Crioula. Under controlled conditions, antibiosis （i.e., reduced longevity, fecundity and increased mortality of the aphid） was observed mainly on the resistant alfalfa cultivars, except on Lahontan. Crioula seemed to be tolerant to aphids. Present data support geographic limitation usage of cultivars, and we suggest Baker and Mesa-Sirsa as sources of antibiosis, and provide biological information of a tropical T. maculata biotype on alfalfa.