Expressing an (E)-β-farnesene synthase in the chloroplast of tobacco affects the preference of green peach aphid and its parasitoid

(E)-β-Famesene(EβF) synthase catalyses the production of EβF,which for many aphids is the main or only component of the alarm pheromone causing the repellence of aphids and also functions as a kairomone for aphids' natural enemies.Many plants possess EβF synthase genes and can release EβF to repel aphids.In order to effectively recruit the plant-derived EβF synthase genes for aphid control,by using chloroplast transit peptide(CTP) of the small subunit of Rubisco(rbcS) from wheat(Triticum aestivum L.),we targeted AαβFS1,an EβF synthase gene from sweet wormwood(Artemisia annua L),to the chloroplast of tobacco to generate CTP + AαβFS1 transgenic lines.The CTP +AαβFS1 transgenic tobacco plants could emit EβF at a level up to 19.25 ng/day per g fresh tissues,4-12 fold higher than the AαβFS1 transgenic lines without chloroplast targeting.Furthermore,aphid/parasitoid behavioral bioassays demonstrated that the CTP + AαβFS1 transgenic tobacco showed enhanced repellence to green peach aphid(Myzus persicae) and attracted response of its parasitoid Diaeretiella rapae,thus affecting aphid infestation at two trophic levels.These data suggest that the chloroplast is an ideal subcellular compartment for metabolic engineering of plant-derived EβF synthase genes to generate a novel type of transgenic plant emitting an alarm pheromone for aphid control.     

Mechanisms for flowering plants to benefit arthropod natural enemies of insect pests： Prospects for enhanced use in agriculture

Reduction of noncrop habitats, intensive use of pesticides and high levels of disturbance associated with intensive crop production simplify the farming landscape and bring about a sharp decline of biodiversity. This, in turn, weakens the biological control ecosystem service provided by arthropod natural enemies. Strategic use of flowering plants to enhance plant biodiversity in a well-targeted manner can provide natural enemies with food sources and shelter to improve biological control and reduce dependence on chemical pesticides. This article reviews the nutritional value of various types of plant-derived food for natural enemies, possible adverse effects on pest management, and the practical application of flowering plants in orchards, vegetables and field crops, agricultural systems where most research has taken place. Prospects for more effective use of flowering plants to maximize biological control of insect pests in agroecosystem are good but depend up on selection of optimal plant species based on information on the ecological mechanisms by which natural enemies are selectively favored over pest species.     

Aphid populations showing differential levels of virulence on Capsicum accessions

The green peach aphid,Myzus persicae,is one of the most threatening pests in pepper cultivation and growers would benefit from resistant varietices.Previously,we identified two Capsicum acessions as susceptible and three as resistant to M.persicae using an aphid population originating from the Netherlands(NL).Later on we identified an aphid population originating from a diferent gcographical region(Switserland,SW)that was virulent on all tested Capsicum acessions.The objeetive of the current work is to describe in detail diferent aspects of the interaction between two aphid populations and two sclected Capsicum acessions(one that was susceptible[PB2013046]and one that was resistant[PB2013071]to population NL),including biochemical processes involved.Electrical penetration graph(EPG)recordings showed similar feeding activities for both aphid populations on PB2013046.On acession PB2013071 the aphid population sw was able to devote significantly more time to phloem ingestion than population NL.We also studied plant defense response and found that plants of acession PB2013046 could not induce an accumulation of reactive oxygen species and callose formation after infestation with either aphid population.However,plants of PB2013071 induced a stronger defense response after infestation by population NL than after infestation by population SW.Based on these results,population SW of M.persicae seems to have overcome the resistance of PB2013071 that prevented feeding of aphids from NL population.The potential mechanism by which SW population overcomes the resistance is discussed.     

Cascade effects of crop species richness on the diversity of pest insects and their natural enemies

Understanding how plant species richness influences the diversity of herbivorous and predatory/parasitic arthropods is central to community ecology.We explore the effects of crop species richness on the diversity of pest insects and their natural enemies.Using data from a four-year experiment with five levels of crop species richness,we found that crop species richness significantly affected the pest species richness,but there were no significant effects on richness of the pests’natural enemies.In contrast,the species richness of pest insects significantly affected their natural enemies.These findings suggest a cascade effect where trophic interactions are strong between adjacent trophic levels,while the interactions between connected but nonadjacent trophic levels are weakened by the intermediate trophic level.High crop species richness resulted in a more stable arthropod community compared with communities in monoculture crops.Our results highlight the complicated cross-trophic interactions and the crucial role of crop diversity in the food webs of agro-ecosystems.     

Plant phenolics mediated bottom-up effects of elevated CO2 on Acyrthosiphon pisum and its parasitoid Aphidius avenae

Elevated concentrations of atmospheric CO2 can alter plant secondary metabolites,which play important roles in the interactions among plants,herbivorous insects and natural enemies.However,few studies have examined the cascading effects of host plant secondary metabolites on tri-trophic interactions under elevated CO2(eCO2).In this study,we determined the effects of eCO2 on the growth and foliar phenolics of Medicago truncatula and the cascading effects on two color genotypes oiAcyrthosiphon pisum(pink vs.green)and their parasitoid Aphidius avenae in the field open-top chambers.Our results showed that eCO2 increased photosynthetic rate,nodule number,yield and the total phenolic content of M.truncatula.eCO2 had contrasting effects on two genotypes of A.pisum;the green genotype demonstrated increased population abundance,fecundity,growth and feeding efficiency,while the pink genotype showed decreased fitness and these were closely associated with the foliar genstein content.Furthermore,eCO2 decreased the parasitic rate of A.avenae independent of aphid genotypes.eCO2 prolonged the emergence time and reduced the emergence rate and percentage of females when associated with the green genotype,but little difference,except for increased percentage of females,was observed in A.avenae under eCO2 when associated with the pink genotype,indicating that parasitoids can perceive and discriminate the qualities of aphid hosts.We concluded that eCO2 altered plant phenolics and thus the performance of aphids and parasitoids.Our results indicate that plant phenolics vary by different abiotic and biotic stimuli and could potentially deliver the cascading effects of eCO2 to the higher trophic levels.Our results also suggest that the green genotype is expected to perform better in future eCO2 because of decreased plant resistance after its infestation and decreased parasitic rate.     

Jasmonate- and salicylate-induced defenses in wheat affect host preference and probing behavior but not performance of the grain aphid,Sitobion avenae

Jasmonate and salicylatemediated signaling pathways play significant roles in induced plant defenses, but there is no sufficient evidence for their roles in monocots against aphids. We exogenously applied methyl jasmonate （MeJA） and salicylic acid （SA） on wheat seedlings and examined biochemical responses in wheat and effects on the grain aphid, Sitobion avenae （Fab.）. Application of MeJA significantly increased levels of wheat＇s polyphenol oxidase, peroxidase and proteinase inhibitor 1, 2 and 6 days after treatment. In twochoice tests, adult aphids preferred control wheat leaves to MeJA or SA treated leaves. Electrical penetration graph （EPG） recordings of aphid probing behavior revealed that on MeJAtreated plants, the duration of aphid＇s first probe was significantly shorter and number of probes was significantly higher than those on control plants. Also total duration of probing on MeJAtreated plants was significantly shorter than on control plants. Total duration of salivation period on SAtreated plants was significantly longer, while mean phloem ingestion period was significantly shorter than on control plants. However, no significant difference in total duration of phloem sap ingestion period was observed among treatments. The EPG data suggest that MeJAdependent resistance factors might be due to feeding deterrents in mesophyll, whereas the SAmediated resistance may be phloembased. We did not observe any significant difference of MeJA and SA application on aphid development, daily fecundity, intrinsic growth rate and population growth. The results indicate that both MeJA and SAinduced defenses in wheat deterred S. avenae colonization processes and feeding behavior, but had no significant effects on its performance.     

Relative importance of long‐term changes in climate and land‐use on the phenology and abundance of legume crop specialist and generalist aphids

Abstract Insect populations are prone to respond to global changes through shifts in phenology, distribution and abundance. However, global changes cover several factors such as climate and land-use, the relative importance of these being largely unknown. Here, we aim at disentangling the effects of climate, land-use, and geographical drivers on aphid abundance and phenology in France, at a regional scale and over the last 40 years. We used aerial data obtained from suction traps between 1978 and 2015 on five aphid species varying in their degree of specialization to legumes, along with climate, legume crop area and geographical data. Effects of environmental and geographical variables on aphid annual abundance and spring migration dates were analyzed using generalized linear mixed models. We found that within the last four decades, aphids have advanced their spring migration by a month, mostly due to the increase in temperature early in the year, and their abundance decreased by half on average, presumably in response to a combination of factors. The influence of legume crop area decreased with the degree of specialization of the aphid species to such crops. The effect of geographical variation was high even when controlling for environmental variables, suggesting that many other spatially structured processes act on aphid population characteristics. Multifactorial analyses helped to partition the effects of different global change drivers. Climate and land-use changes have strong effects on aphid populations, with important implications for future agriculture. Additionally, trait-based response variation could have major consequences at the community scale.     

Influence of cornicle droplet secretions of the cabbage aphid,Brevicoryne brassicae,on parasitism behavior of na＇fve and experienced Diaeretiella rapae

Insects have evolved amazing methods of defense to ward off enemies. Many aphids release cornicle secretions when attacked by predators and parasitoids. These se cretions contain an alarm pheromone that alerts other colony members of danger, thereby providing indirect fitness benefits to the releaser. In addition, contact with cornicle se cretions could also threaten an attacker and could provide direct fitness to the releaser. However, cornicle secretions may also be recruited as a kairomonal cue by aphid natural enemies. In this study, we investigated the effect of the cornicle droplet volatiles of the cabbage aphid, Brevicoryne brassicae （L.）, on the hostsearching behavior of naive and experienced female Diaeretiella rapae （M＇Intosh） parasitoids in olfactometer studies. In addition, we evaluated the role ofB. brassicae cornicle droplets on the oviposition prefer ence of the parasitoid in a twochoice bioassay. Naive females did not exhibit any preference between volatiles from aphids secreting cornicle droplets over nonsecreting aphids, while experienced parasitoids exploited the secretions in their host location. Experienced females were also able to choose volatiles from both secreting and nonsecreting aphids over clean air, while this ability was not observed in naive females. Although secretion of cornicle droplets did not influence the percentage of first attack in either naive or experienced females, the success of attack （i.e. resulting in a larva） was significantly different between secreting and nonsecreting aphids in the case of experienced parasitoids.     

How does the host-specialized aphid deal with food deficiency？

Aphis gossypii Glover shows obvious host specialization, with cucurbit- and cotton-specialized biotypes or host races in many regions. Because its annual natal hostcrops senesce earlier the cucurbit-specialized biotype may suffer food deficiency. The method this biotype uses to overcome this challenge is still poorly understood. In orderto understand the potential of the cucurbit-specialized biotype aphids in host shift and usage, the performance of this biotype on cotton （Gossypium hirsutum）, a common butpoor quality host plant, was explored in this study. The cucurbit-specialized aphids could establish populations on cotton only when these plants had at least nine leaves, and subsequent populations developed rather slowly. The presence of whitefly populations on cotton improved the success rate of cucurbit-specialized aphids. The cucurbit-specialized aphidswere mainly distributed on the older leaves of cotton, with only a few settling on the upper leaves. The cucurbit-specialized aphids reared on cotton for 40, 54 and 61 days stillmaintained strong preference for their natal host plant, cucumber （Cucumis sativus）, rather than cotton, and their net reproductive rates and intrinsic rates of natural increase weredramatically lower when they were transferred onto new six-leaf cotton plants or detached leaves. Therefore, we concluded that the cucurbit-specialized aphids have the potentialto utilize mature or whitefly-stressed cotton plants, but that this feeding experience on cotton did not alter their specialization for cucurbits. Some cotton plants could act as atemporary host for the cucurbit-specialized aphids to overcome food deficiency arising from senescing cucurbits.     

表达尾穗苋凝集素基因的转基因烟草对桃蚜(Myzus persicae)繁殖的影响(英文)

To investigate the possible function of the agglutinin from Amaranthus caudatus L. (ACA) in plant defending against insect pests, ACA cDNA was cloned by RT-PCR and the 5‘ and 3‘ sequences were confirmed by rapid amplification of cDNA ends (RACE). The phloem-specific expression vector of ACA gene, pBCACAc, was constructed based on the plant binary vector pBC438 and transfered into tobacco plants via Agrobacterium-mediated transformation method. Results from PCR and Southern blotting analysis showed that AOA gene was integrated into the genomes of transformed plants and the transgene integration varied from one to four estimated copies per genome. Western blotting analysis indicated that ACA gene was transcribed and translated in the transgenic plants. The bioassay of Myzus persicae Sulzer on detached leaves demonstrated that the 78% transgenic tobacco plants displayed an average aphid-resistant rate of more than 75%. Some apterous progeny of M. persicae were found dead on the resistant plants. These results indicate that ACA gene should be an effective aphid-resistant gene and could be valuable for application in crop breeding for aphid resistance.     

Response of Solanum tuberosum to Myzus persicae infestation at different stages of foliage maturity

Young leaves of the potato Solanum tuberosum L. cultivar Kardal contain resistance factors to the green peach aphid Myzuspersicae （Sulzer） （Hemiptera： Aphididae） and normal probing behavior is impeded. However, M. persicae can survive and reproduce on mature and senescent leaves of the cv. Kardal plant without problems. We compared the settling ofM. persicae on young and old leaves and analyzed the impact of aphids settling on the plant in terms of gene expression. Settling, as measured by aphid numbers staying on young or old leaves, showed that after 21 h significantly fewer aphids were found on the young leaves. At earlier time points there were no difference between young and old leaves, suggesting that the young leaf resistance factors are not located at the surface level but deeper in the tissue. Gene expression was measured in plants at 96 h postinfestation, which is at a late stage in the interaction and in compatible interactions this is long enough for host plant acceptance to occur. In old leaves of cv. Kardal （compatible interaction）, M. persicae infestation elicited a higher number of differentially regulated genes than in young leaves. The plant response to aphid infestation included a larger number of genes induced than repressed, and the proportion of induced versus repressed genes was larger in young than in old leaves. Several genes changing expression seem to be involved in changing the metabolic state of the leaf from source to sink.     

Sieve element occlusion provides resistance against Aphis gossypii in TGR-1551 melons

Feeding behavior and plant response to feeding were studied for the aphid Aphis gossypii Glover on susceptible and resistant melons(cv.Iroquois and TGR-1551,respectively).Average phloem phase bout duration on TGR-1551 was<7% of the duration on Iroquois.Sixty-seven percent of aphids on TGR-1551 never produced a phloem phase that attained ingestion(EPG waveform E2)in contrast to only 7% of aphids on Iroquois.Average bout duration of waveform E2(scored as zero if phloem phase did not attain E2)on TGR-1551 was<3% of the duration on Iroquois.Conversely,average bout duration of EPG waveform El(sieve element salivation)was 2.8 times greater on TGR-1551 than on Iroquois.In a second experiment,liquid nitrogen was used to rapidly cryofix leaves and aphids within a few minutes after the aphids penetrated a sieve element.Phloem near the penetration site was then examined by confocal laser scanning microscopy.Ninety-six percent of penetrated sieve elements were occluded by protein in TGR-1551 in contrast to only 28% in Iroquois.Usually in TGR-1551,occlusion was also observed in nearby nonpenetrated sieve elements.Next,a calcium channel blocker,trivalent lanthanum,was used to prevent phloem occlusion in TGR-1551,and A.gossypii feeding behavior and the plants phloem response were compared between lanthanum-treated and control TGR-1551.Lanthanum treatment eliminated the sieve element protein occlusion response and the aphids readily ingested phloem sap from treated plants.This study provides strong evidence that phloem occlusion is a mechanism for resistance against A.gossypii in TGR-1551.     

The price of protection:a defensive endosymbiont impairs nymph growth in the bird cherry-oat aphid,Rhopalosiphum padi

Bacterial endosymbionts have enabled aphids to adapt to a range of stressors,but their effects in many aphid species remain to be established.The bird cherry-oat aphid,Rhopalosiphum padi(Linnaeus),is an important pest of cereals worldwide and has been reported to form symbiotic associations with Serratia symbiotica and Sitobion miscanthi L-type symbiont endobacteria,although the resulting aphid phenotype has not been described.This study presents the first report of R.padi infection with the facultative bacterial endosymbiont Hamiltonella defensa.Individuals of R.padi were sampled from populations in Eastern Scotland,UK,and shown to represent seven R.padi genotypes based on the size of polymorphic microsatellite markers;two of these genotypes harbored H.defensa.In parasitism assays,survival of H.defensa-infected nymphs following attack by the parasitoid wasp Aphidius colemani(Viereck)was 5 fold higher than for uninfected nymphs.Aphid genotype was a major determinant of aphid performance on two Hordeum species,a modern cultivar of barley H.vulgare and a wild relative H.spontaneum,although aphids infected with H.defensa showed 16%lower nymph mass gain on the partially resistant wild relative compared with uninfected individuals.These findings suggest that deploying resistance traits in barley will favor the fittest R.padi genotypes,but symbiontinfected individuals will be favored when parasitoids are abundant,although these aphids will not achieve optimal performance on a poor quality host plant.     

Creation of Transgenic Bananas Expressing Human Lysozyme Gene for Panama Wilt Resistance

Human lysozyme （HL） inhibits Fusarium oxysporum （FocR4） growth in vitro. To obtain transgenic bananas （Musa spp.） that are resistant to Panama wilt （F. oxysporum）, we introduced an HL gene that is driven by a constitutive cauliflower mosaic virus 35S promoter into the banana via Agrobacteriummediated transformation. PCR confirmed that 51 transgenic plants were obtained. The development of Panama wilt symptoms were examined after the plants had been grown in pots. The non-transgenic plants developed typical fusarium symptoms 60 d after FocR4 inoculation, whereas 24 of 51 transgenic plants remained healthy. The transgenic banana plants that showed resistance to FocR4 in the pots were then planted in a field that was heavily infected with FocR4 for further investigation. Eleven of 24 plants devel- oped symptoms before bud emergence; another 11 plants showed symptoms after bud emergence and the remaining two plants, H-67 and H-144, remained healthy and were able to fruit. Northern blotting analysis demonstrated that H-67 and H-144, bearing the strongest resistance to Panama wilt, had the highest level of HL expression and that the expression of HL was well correlated with the FocR4 resistance of transgenic plants. We conclude that Agrobacterium-mediated transformation, with the assistance of particle bombardment, is a powerful approach for banana transformation and that a transgenic HL gene can cause resistance of the crop to FocR4 in the field.     

Diversity of bacteria associated with Hormaphidinae aphids (Hemiptera: Aphididae)

Bacteria are ubiquitous inhabitants of animals.Hormaphidinae is a particular aphid group exhibiting very diverse life history traits.However,the microbiota in this group is poorly known.In the present study,using high-throughput sequencing of bacterial 16S ribosomal RNA gene amplicons,we surveyed the bacterial flora in hormaphidine aphids and explored whether the aphid tribe,host plant and geographical distribution are associated with the distribution of secondary symbionts.The most dominant bacteria detected in hormaphidine species are heritable symbionts.As expected,the primary endosymbiont Buchnera aphidicola is the most abundant symbiont across all species and has cospeciated with its host aphids.Six secondary symbionts were detected in Hormaphidinae.Arsenophonus is widespread in Hormaphidinae species,suggesting the possibility of ancient acquisition of this symbiont.Ordination analyses and statistical tests show that the symbiont composition does not seem to relate to any of the aphid tribes,host plants or geographical distributions,which indicate that horizontal transfers might occur for these symbionts in Hormaphidinae.Correlation analysis exhibits negative interference between Buchnera and coexisting secondary symbionts,while the interactions between different secondary symbionts are complicated.These findings display a comprehensive picture of the microbiota in Hormaphidinae and may be helpful in understanding the symbiont diversity within a group of aphids.     

Inheritance patterns of secondary symbionts during sexual reproduction of pea aphid biotypes

Herbivorous insects frequently harbor bacterial symbionts that affect their ecol- ogy and evolution. Aphids host the obligatory endosymbiont Buchnera, which is requiredfor reproduction, together with facultative symbionts whose frequencies vary across aphid populations. These maternally transmitted secondary symbionts have been particularlystudied in the pea aphid, Acyrthosiphon pisum, which harbors at least 8 distinct bacterial species （not counting Buchnera） having environmentally dependent effects on host fitness.In particular, these symbiont species are associated with pea aphid populations feeding on specific plants. Although they are maternally inherited, these bacteria are occasionallytransferred across insect lineages. One mechanism of such nonmaternal transfer is paternal transmission to the progeny during sexual reproduction. To date, transmission of secondarysymbionts during sexual reproduction of aphids has been investigated in only a handful of aphid lineages and 3 symbiont species. To better characterize this process, we investigatedinheritance patterns of 7 symbiont species during sexual reproduction of pea aphids through a crossing experiment involving 49 clones belonging to 9 host-specialized biotypes, and117 crosses. Symbiont species in the progeny were detected with diagnostic qualitative PCR at the fundatrix stage hatching from eggs and in later parthenogenetic generations.We found no confirmed case of paternal transmission of symbionts to the progeny, and we observed that maternal transmission of a particular symbiont species （Serratia symbiotica）was quite inefficient. We discuss these observations in respect to the ecology of the pea aphid.     

Potassium and phosphorus transport and signaling in plants

Nitrogen(N), potassium(K), and phosphorus(P) are essential macronutrients for plant growth and development, and their availability affects crop yield. Compared with N, the relatively low availability of K and P in soils limits crop production and thus threatens food security and agricultural sustainability. Improvement of plant nutrient utilization efficiency provides a potential route to overcome the effects of K and P deficiencies. Investigation of the molecular mechanisms underlying how plants sense, absorb, transport, and use K and P is an important prerequisite to improve crop nutrient utilization efficiency. In this review, we summarize current understanding of K and P transport and signaling in plants, mainly taking Arabidopsis thaliana and rice(Oryza sativa) as examples. We also discuss the mechanisms coordinating transport of N and K, as well as P and N.     

Effect of plant nutrition on aphid size,prey consumption,and life history characteristics of green lacewing

Plant quality can directly and indirectly affect the third trophic level. The predation by all the instars of green lacewing, Chrysoperla carnea （S.） （Neuroptera： Chrysopidae） on the cereal aphids, Rhopalosiphum padi （L.）, and Sitobion avenae （F.） at varying nitrogen fertilizer levels was calculated under laboratory conditions. Wheat plants were grown on four nitrogen fertilizer levels and aphids were fed on these plants and subsequently offered as food to the C. carnea. Aphid densities of 10, 30, and 90 were offered to first, second, and third instar larvae of green lacewing. Increased nitrogen application improved nitrogen contents of the plants and also the body weight of cereal aphids feeding on them. Aphid consumption by green lacewings was reduced with the increase in nitrogen content in the host plants of aphids. Predation of both aphid species by first, second, and third instars larvae of C. carnea was highest on aphids reared on plants with the lowest rate of fertilization, suggesting a compensatory consumption to overcome reduced biomass （lower aphid size）. Total biomass devoured by C. carnea on all nitrogen fertilizer treatments was not statistically different. Additionally, the heavier host prey influenced by the plant nutrition had an effect on the life history characteristics of green lacewings. The larval duration, pupal weight, pupal duration, fecundity, and male and female longevity were significantly affected by the level of nitrogen fertilization to the aphid＇s host plants, except for pupal duration when fed on S. avenae. This study showed that quantity of prey supplied to the larvae affects the prey consumption and thereafter the life history characteristics of green lacewings.     

Aphidophagous guilds on nettle （Urtica dioica） strips close to fields of green pea, rape and wheat

The common nettle （Urtica dioica L.） is a perennial and cosmopolitan plant species and is known to be the source of food for a great diversity of insects. To understand the importance of the nettle in agro-ecosystems, a field experiment was carried out in an experimental farm at Gembloux （Belgium） to study the effect of nettle margin strips on aphid and aphidophagous populations in close field crops, namely wheat （Triticum aestivum L.）, green pea （Pisum sativum L.） and rape （Brassicae napus L.）. The aphids and related beneficial populations were weekly assessed, from March to August 2005, by visual observations in two plots per field crop. A higher abundance of aphidophagous beneficials was collected in nettle strips when compared to the field crops. Particularly, the presence of predatory anthocorids, mirids and green lacewings was observed on nettle only. Nevertheless, the most abundant aphid predatory family, the Coccinellidae, was distributed in both environments, in nettle strips and in crop fields. The field margin supported a significantly higher density ofHarmonia axyridis than the field crops. In contrast, the field crops, green pea particularly, supported a higher density of Coccinella septempunctata. The distribution of the aphidophagous species, mainly the ladybirds, was discussed in relation to the host plant and related aphid species and their potential effect on integrated pest management.     

Progress in TILLING as a tool for functional genomics and improvement of crops

Food security is a global concern and substantial yield increases in crops are required to feed the growing world population. Mutagenesis is an important tool in crop improvement and is free of the regulatory restrictions imposed on genetically modified organisms. Targeting Induced Local Lesions in Genomes（TILLING）, which combines traditional chemical mutagenesis with high‐throughput genome‐wide screening for point mutations in desired genes, offers a powerful way to create novel mutant alleles for both functional genomics and improvement of crops. TILLING is generally applicable to genomes whether small or large, diploid or evenallohexaploid, and shows great potential to address the major challenge of linking sequence information to the function of genes and to modulate key traits for plant breeding. TILLING has been successfully applied in many crop species and recent progress in TILLING is summarized below, especially on the developments in mutation detection technology, application of TILLING in gene functional studies and crop breeding. The potential of TILLING/EcoTILLING for functional genetics and crop improvement is also discussed. Furthermore, a small‐scale forward strategy including backcross and selfing was conducted to release the potential mutant phenotypes masked in M2（or M3） plants.